Macrocyclic inhibitors of flaviviridae viruses

ABSTRACT

Provided are compounds of Formula I: 
     
       
         
         
             
             
         
       
     
     and pharmaceutically acceptable salts and esters thereof. The compounds, compositions, and methods provided are useful for the treatment of virus infections, particularly hepatitis C infections.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §119(e) to U.S.Provisional Application Ser. No. 61/657,562, filed on Jun. 8, 2012, theentirety of which is incorporated herein by reference.

FIELD

The present application provides novel compounds inhibiting viruses,compositions containing such compounds, and therapeutic methodscomprising the administration of such compounds.

BACKGROUND

RNA viruses comprising the Flaviviridae family include at least threedistinguishable genera including pestiviruses, flaviviruses, andhepaciviruses (Calisher, et al., J. Gen. Virol., 1993, 70, 37-43). Whilepestiviruses cause many economically important animal diseases such asbovine viral diarrhea virus (BVDV), classical swine fever virus (CSFV,hog cholera) and border disease of sheep (BDV), their importance inhuman disease is less well characterized (Moennig, V., et al., Adv. Vir.Res. 1992, 48, 53-98). Flaviviruses are responsible for important humandiseases such as dengue fever and yellow fever while hepaciviruses causehepatitis C virus infections in humans. Other important viral infectionscaused by the Flaviviridae family include West Nile virus (WNV) Japaneseencephalitis virus (JEV), tick-borne encephalitis virus, Junjin virus,Murray Valley encephalitis, St Louis enchaplitis, Omsk hemorrhagic fevervirus and Zika virus.

The hepatitis C virus (HCV) is the leading cause of chronic liverdisease worldwide (Boyer, N. et al. J Hepatol. 32:98-112, 2000) so asignificant focus of current antiviral research is directed toward thedevelopment of improved methods of treatment of chronic HCV infectionsin humans (Di Besceglie, A. M. and Bacon, B. R., Scientific American,Oct.: 80-85, (1999); Gordon, C. P., et al., J. Med. Chem. 2005, 48,1-20; Maradpour, D., et al., Nat. Rev. Micro. 2007, 5(6), 453-463). Anumber of HCV treatments are reviewed by Dymock et al. in AntiviralChemistry & Chemotherapy, 11:2; 79-95 (2000). Virologic cures ofpatients with chronic HCV infection are difficult to achieve because ofthe prodigious amount of daily virus production in chronically infectedpatients and the high spontaneous mutability of HCV virus (Neumann, etal., Science 1998, 282, 103-7; Fukimoto, et al., Hepatology, 1996, 24,1351-4; Domingo, et al., Gene 1985, 40, 1-8; Martell, et al., J. Virol.1992, 66, 3225-9.

Currently, there are primarily two antiviral compounds, ribavirin, anucleoside analog, and interferon-alpha (α) (IFN), that are used for thetreatment of chronic HCV infections in humans. Ribavirin alone is noteffective in reducing viral RNA levels, has significant toxicity, and isknown to induce anemia. The combination of IFN and ribavirin has beenreported to be effective in the management of chronic hepatitis C(Scott, L. J., et al. Drugs 2002, 62, 507-556) but less than half thepatients given this treatment show a persistent benefit. Therefore,there is a need to develop more effective anti-HCV therapies.

The macrocycle sanglifehrin and derivatives are immunomodulatory andbind peptidyl-prolyl cis/trans isomerase (PPlase) cyclophilins in aunique manner (WO 97/02285; WO 98/07743; J. Am. Chem. Soc 2003, 125,3849-3859; J. Org. Chem. 2000, 65, 9255-9260; Angew. Chem. Int. Ed.1999, 38, 2443-2446). The cyclophilins are peptidyl-prolyl cis/transisomerases (PPlase) that regulate protein folding in vivo and inhibithepatitis C virus (Lin et al., WO2006/138507). However, none of thesanglifehrins or their derivatives has become available for humananti-viral therapy. Therefore, there is a continuing need to developmacrocyclic sanglifehrins with anti-Flaviviridae virus activity andparticularly anti-HCV activity.

SUMMARY

In one embodiment, there is provided a compound represented by FormulaI:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein:

A is a bond, —O—, —S(O)_(n)—, —NH—, —N((C₁-C₄)alkyl)- or(C₁-C₂)alkylene;

A¹ is (C₁-C₈)alkylene, (C₂-C₅)alkenylene, (C₂-C₅)alkynylene, arylene,heteroarylene, cycloalkylene, heterocycloalkylene, aryl(C₁-C₂)alkylene,heteroaryl(C₁-C₂)alkylene, cycloalkyl(C₁-C₂)alkylene orheterocycloalkyl(C₁-C₂)alkylene, wherein a sp³ carbon atom of A¹ isoptionally replaced by —O—, —S(O)_(n)—, —NH— or —N((C₁-C₄)alkyl)-, andwherein a sp³ or sp² carbon atom of A¹ is optionally substituted withone or more substituents selected from the group consisting of halo,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, aryl, heterocycloalkyl,cycloalkyl, aryl(C₁-C₄)alkyl, cycloalkyl(C₁-C₄)alkyl,heterocycloalkyl(C₁-C₄)alkyl, arylheterocycloalkyl(C₁-C₄)alkyl, —OR⁹,—SR⁹, —S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂ where such an optional substitutionis chemically feasible;

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)— cycloalkylene, arylene,cycloalkylene, (C₁-C₃)alkylene, (C₂-C₃)alkenylene or (C₂-C₃)alkynylene,wherein the arylene, heteroarylene, heterocycloalkylene or cycloalkylenemoiety of A² is optionally substituted with one or more substituentsselected from the group consisting of —OR⁹, —SR⁹, —S(O)R⁹, —S(O)₂R⁹,—N(R⁹)₂, halo, halo(C₁-C₄)alkyl, halo(C₁-C₄)alkoxy cyano and(C₁-C₈)alkyl where such an optional substitution is chemically feasible;

L¹ is —O—C(O)—, —O—CH₂—, —NR¹¹—C(O)—, —NR¹⁰—CH₂—, —NH—C(R¹⁰)₂— or—NH—S(O)₂—, wherein each R¹⁰ is independently H, (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, (C₁-C₄)alkenyl or cycloalkyl; and R¹¹ is (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, (C₁-C₄)alkenyl or cycloalkyl;

X¹ is a bond, —O—, —NH—, —N((C₁-C₄)alkyl)- or heterocycloalkylene;

R¹ and R² are independently H, (C₁-C₄)alkyl, (C₂-C₄)alkenyl,(C₂-C₄)alkynyl, halo, cyano or —C(O)—(C₁-C₄)alkyl; or

R¹ and R², when taken together with the carbon to which they are bothattached, form —C(═O)—, —C(═S)— or —C(═N(C₁-C₄)alkyl)-;

R³ is H or (C₁-C₄)alkyl which is optionally substituted with halo,cyano, hydroxyl or (C₁-C₄)alkoxy;

R^(4a) and R^(4b) are independently H, (C₁-C₈)alkyl, aryl,aryl(C₁-C₄)alkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl,cycloalkyl or cycloalkyl(C₁-C₄)alkyl, wherein each R^(4a) and R^(4b) isoptionally substituted with one or more substituents selected from thegroup consisting of cyano, —COOH, halo, hydroxyl, (C₁-C₈)alkoxy, amino,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, aryl and heteroaryl wheresuch an optional substitution is chemically feasible;

R^(5a) and R^(5b) are independently H, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, (C₁-C₈)alkoxy, aryl, heterocycloalkyl, cycloalkyl,aryl(C₁-C₄)alkyl, cycloalkyl(C₁-C₄)alkyl orheterocycloalkyl(C₁-C₄)alkyl, wherein each R^(5a) and R^(5b) isoptionally substituted with one or more substituents selected from thegroup consisting of —N₃, cyano, —COOH, halo, hydroxyl, amino,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, (C₁-C₈)alkoxy, aryl andheteroaryl, or

R^(5a) and R^(5b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹;

R^(6a) and R^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy, aryl, cycloalkyl, heterocycloalkyl,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹, —NHC(O)R⁹ and(C₁-C₈)alkanoyl where such an optional substitution is chemicallyfeasible; or R^(6a) and R^(6b) together form a spirocycle having Formula(a);

each R⁸ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl, (C₂-C₄)alkynyl, aryl, heteroaryl, heterocycloalkyl orcycloalkyl, wherein R⁸ is optionally substituted with —OR, —N(R⁹)₂,—CON(R⁹)₂ or cyano;

each R⁹ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl or (C₂-C₄)alkynyl;

each n is independently 0, 1 or 2; and

m is 1, 2, 3, 4 or 5.

In another embodiment, there is provided a pharmaceutical compositioncomprising a compound of Formula I or a pharmaceutically acceptablesalt, isotope, stereoisomer, mixture of stereoisomers, tautomer, esteror prodrug thereof and one or more pharmaceutically acceptable carriersor excipients. In one aspect of the embodiment, the pharmaceuticalcomposition further comprises one or more additional therapeutic agents.

In yet another embodiment, a method for treating Flaviviridae viralinfection is provided comprising administering a therapeuticallyeffective amount of a compound of Formula I or a pharmaceuticallyacceptable salt, isotope, stereoisomer, mixture of stereoisomers,tautomer, ester or prodrug thereof to a mammal in need thereof. In oneaspect of the embodiment, the treatment results in the reduction of thein viral load or clearance of viral RNA in a patient.

In yet another embodiment, a method for treating Coronaviridae viralinfection is provided comprising administering a therapeuticallyeffective amount of a compound of Formula I or a pharmaceuticallyacceptable salt, isotope, stereoisomer, mixture of stereoisomers,tautomer, ester or prodrug thereof to a mammal in need thereof. In oneaspect of the embodiment, the treatment results in the reduction of thein viral load or clearance of viral RNA in a patient.

DETAILED DESCRIPTION Definitions

Unless stated otherwise, the following terms and phrases as used hereinare intended to have the following meanings:

“Alkanoyl” is RC(O)—; “alkanoyloxy” is RC(O)O—; and “alkanoylamino” isRC(O)NR′—; where R is an alkyl group as defined herein, and R¹ ishydrogen or alkyl.

“Alkenyl” refers to a straight or branched hydrocarbyl group with atleast one site of unsaturation, i.e., a carbon-carbon, sp² double bond.In some embodiments, alkenyl is a C₂—O₂₀ alkenyl group, a C₂-C₁₀ alkenylgroup or a C₂-C₆ alkenyl group. Examples of alkenyl group include, butare not limited to, vinyl (—CH═CH₂), allyl (—CH₂CH═CH₂), cyclopentenyl(—C₅H₇), and 5-hexenyl (—CH₂CH₂CH₂CH₂CH═CH₂).

“Alkenylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical having two monovalent radical centers derivedby the removal of two hydrogen atoms from the same or two differentcarbon atoms of a parent alkene. For example, and alkenylene group canhave 2 to 20 carbon atoms, 2 to 10 carbon atoms, or 2 to 6 carbon atoms.Typical alkenylene radicals include, but are not limited to,1,2-ethenylene (—CH═CH—).

“Alkoxy” is RO— where R is alkyl, as defined herein. Non-limitingexamples of alkoxy groups include methoxy, ethoxy and propoxy.

“Alkyl” refers to a straight or branched chain hydrocarbyl group. In anembodiment, alkyl has from 1 to 20 carbon atoms (i.e., C₁-C₂₀ alkyl). Insome embodiments, alkyl is a C1-C₁₀ alkyl group or a C₁-C₆ alkyl group.Examples of alkyl groups include, but are not limited to, methyl, ethyl,propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl,octyl, nonyl and decyl.

“Alkylene” refers to a saturated, branched or straight chain radical orcyclic hydrocarbon radical having two monovalent radical centers derivedby the removal of two hydrogen atoms from the same or two differentcarbon atoms of a parent alkane. For example, an alkylene group can have1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms.Examples of alkylene radicals include, but are not limited to, methylene(—CH₂—), ethylene (—CH₂CH₂—), propylene (—CH₂CH₂CH₂—) and butylene(—CH₂CH₂CH₂CH₂—).

“Alkynyl” refers to a hydrocarbon containing normal, secondary ortertiary carbon atoms with at least one site of unsaturation, i.e., acarbon-carbon, sp triple bond. For example, an alkynyl group can have 2to 20 carbon atoms (i.e., C₂-C₂₀ alkynyl), 2 to 12 carbon atoms (i.e.,C₂-C₁₂ alkyne,) or 2 to 6 carbon atoms (i.e., C₂-C₆ alkynyl). Examplesof alkynyl groups include, but are not limited to, acetylenic (—C≡CH)and propargyl (—CH₂C≡CH).

“Amino” refers to —NH₂.

“Alkylamino” refers to an amino group substituted with one or more alkylgroups. “Mono(alkyl)amino” or “(alkyl)amino” is RNH—, and“di(alkyl)amino” or “(alkyl)₂-amino” is R₂N—, where each of the R groupsis alkyl as defined herein and are the same or different. Examples ofalkylamino groups include, but are not limited to, methylamino,ethylamino, propylamino, butylamino, dimethylamino, diethylamino andmethylethylamno.

“Alkynylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical having two monovalent radical centers derivedby the removal of two hydrogen atoms from the same or two differentcarbon atoms of a parent alkyne. For example, an alkynylene group canhave 2 to 20 carbon atoms, 2 to 10 carbon atoms, or 2 to 6 carbon atoms.Typical alkynylene radicals include, but are not limited to, acetylene(—C≡C—), propargylene (—CH₂C≡C—), and 4-pentynylene (—CH₂CH₂CH₂C≡C—).

“Aryl” refers to any monocyclic or bicyclic carbon ring of up to 7 atomsin each ring, wherein at least one ring is aromatic, or an aromatic ringsystem of 5 to 14 carbons atoms which includes a carbocyclic aromaticgroup fused with a 5- or 6-membered cycloalkyl group. Examples of arylgroups include, but are not limited to, phenyl, naphthyl,tetrahydronaphthyl and indanyl.

“Arylalkyl” refers to an alkyl as defined herein substituted with anaryl radical.

“Arylene” refers to an aryl as defined above having two monovalentradical centers derived by the removal of two hydrogen atoms from twodifferent carbon atoms of a parent aryl. Typical arylene radicalsinclude, but are not limited to, phenylene, e.g.,

and naphthylene, e.g.,

“Arylalkylene” refers to an arylalkyl as defined above having twomonovalent radical centers derived by the removal of one hydrogen atomfrom the aryl radical and the other hydrogen removed from the alkylradical of the group.

“Cycloalkyl” refers to a hydrocarbyl group containing at least onesaturated or partially unsaturated ring structure, and attached via aring carbon. Cycloalkyl groups include hydrocarbon mono-, bi-, andpoly-cyclic rings, whether fused, bridged, or spiro. In variousembodiments, it refers to a saturated or a partially unsaturated C₃-C₁₂cyclic moiety, examples of which include cyclopropyl, cyclobutyl,cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl andcyclooctyl.

“Cycloalkylalkyl” refers to an alkyl moiety substituted with acycloalkyl group. Examples of cycloalkylalkyl groups includecyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl andcyclohexylmethyl.

“Cycloalkylalkylene” refers to an alkylene moiety substituted with acycloalkyl group. Examples of cycloalkylalkylene groups includecyclopropylmethylene, cyclobutylmethylene, cyclopentylethylene andcyclohexylmethylene.

“Cycloalkylene” refers to a cycloalkyl, as defined herein, having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent cycloalkyl.Examples of cycloalkylene include, but are not limited to,cyclopropylene, cyclobutylene, cyclopentylene and cyclohexylene.

“Dialkylaminoalkyl” refers to an alkyl moiety substituted with adialkylamino group, wherein dialkylamino is as defined herein.

“Ester” means any ester of a compound in which any of the —COOHfunctions of the molecule is replaced by a —C(O)OR function, or in whichany of the —OH functions of the molecule are replaced with a —OC(O)Rfunction, in which the R moiety of the ester is any carbon-containinggroup which forms a stable ester moiety, including but not limited toalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl,heterocyclyl, heterocyclylalkyl and substituted derivatives thereof.

“Halo” refers to chloro (—Cl), bromo (—Br), fluoro (—F) or iodo (—I).

“Haloalkoxy” refers to alkoxy, as defined herein, substituted with oneor more halo radicals.

“Haloalkoxyalkyl” refers to an alkyl moiety substituted with ahaloalkoxy group, as defined herein.

“Haloalkyl” refers to an alkyl group, in which one or more hydrogenatoms of the alkyl group is replaced with a halogen atom. Examples ofhaloalkyl groups include, but are not limited to, —CF₃, —CHF₂, —CFH₂ and—CH₂CF₃.

“Heterocycloalkyl” refers to a saturated or partially unsaturatedmonocyclic, bicyclic or tricyclic group of 2 to 14 ring-carbon atomsand, in addition to ring-carbon atoms, 1 to 4 heteroatoms selected fromthe group consisting of P, N, O and S. The heterocyclic group can beattached through a carbon atom or through a heteroatom, and whensubstituted, the substituent can be bonded to a carbon atom or aheteroatom. Examples of heterocyclyl include azetidinyl,benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl,benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl,cinnolinyl, furanyl, imidazolyl, indolinyl, indolyl, indolazinyl,indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl,isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline,isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl,pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl,quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl,tetrahydrothiopyranyl, tetrahydroisoquinolinyl, tetrazolyl,tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl,azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl,pyridin-2-onyl, pyrrolidinyl, morpholinyl, thiomorpholinyl,dihydrobenzoimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl,dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl,dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl,dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl andN-oxides thereof.

“Arylheterocycloalkyl” refers to a heterocycloalkyl group, as definedherein, in which a hydrogen atom has been replaced with an aryl group.

“Arylheterocycloalkylalkyl” refers to an alkyl group, as defined herein,in which a hydrogen atom has been replaced with a heterocycloalkylgroup, and a hydrogen atom of the heterocycloalkyl group has beenreplaced with an aryl group.

“Heterocycloalkylalkyl” refers to an alkyl group, as defined herein, inwhich a hydrogen atom has been replaced with a heterocycloalkyl group.

“Heterocycloalkylalkylene” refers to an alkylene group, as definedherein, in which a hydrogen atom has been replaced with aheterocycloalkyl group.

“Heterocycloalkylene” refers to a heterocycloalkyl, as defined above,having two monovalent radical centers derived by the removal of twohydrogen atoms from the same or two different carbon atoms of a parentheterocycloalkyl group.

“Heteroaryl” refers to a monocyclic, bicyclic or tricyclic ring havingup to 7 atoms in each ring, wherein at least one ring is aromatic andcontains from 1 to 4 heteroatoms in the ring selected from the groupconsisting of N, O and S, Non-limiting examples of heteroaryl includepyridyl, thienyl, furanyl, pyrimidyl, imidazolyl, pyranyl, pyrazolyl,thiazolyl, thiadiazolyl, isothiazolyl, oxazolyl, isoxazoyl, pyrrolyl,pyridazinyl, pyrazinyl, quinolinyl, isoquinolinyl, benzofuranyl,dibenzofuranyl, dibenzothiophenyl, benzothienyl, indolyl,benzothiazolyl, benzooxazolyl, benzimidazolyl, isoindolyl,benzotriazolyl, purinyl, thianaphthenyl and pyrazinyl. Attachment ofheteroaryl can occur via an aromatic ring, or, if heteroaryl is bicyclicor tricyclic and one of the rings is not aromatic or contains noheteroatoms, through a non-aromatic ring or a ring containing noheteroatoms. “Heteroaryl” is also understood to include the N-oxidederivative of any nitrogen containing heteroaryl.

“Heteroarylalkyl” refers to an alkyl group, as defined herein, in whicha hydrogen atom has been replaced with a heteroaryl group.

“Heteroarylalkylene” refers to an alkylene group, as defined herein, inwhich a hydrogen atom has been replaced with a heteroaryl group.

“Heteroarylene” refers to a heteroaryl, as defined above, having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent heteroarylgroup. Non-limiting examples of heteroarylene groups are:

“Hydroxyalkoxy” refers to an alkoxy, as defined herein, substituted witha hydroxyl group (—OH). An example of hydroxyalkoxy is hydroxyethoxy.

“Hydroxyalkyl” refers to an alkyl group substituted with at least onehydroxyl group. Examples of hydroxyalkyl groups include, but are notlimited to, hydroxymethyl, hydroxyethyl, hydroxypropyl and hydroxybutyl.

“Prodrug” refers to any compound that when administered to a biologicalsystem generates the drug substance, i.e., active ingredient, as aresult of spontaneous chemical reaction(s), enzyme catalyzed chemicalreaction(s), photolysis, and/or metabolic chemical reaction(s). Aprodrug is thus a covalently modified analog or latent form of atherapeutically active compound. Non-limiting examples of prodrugsinclude ester moieties, quaternary ammonium moieties, glycol moieties,and the like.

The term “optionally substituted” refers to a moiety wherein allsubstituents are hydrogen or wherein one or more of the hydrogens of themoiety are replaced by non-hydrogen substituents. Multiple substitutionson the same atom are also permitted where chemically feasible (e.g., adioxo substitution to provide —S(O)₂—, geminal substituents, spirocycloalkyl or heterocycloalkyl rings, etc.). In some embodiments, “oneor more” substituents is from one to three substituents.

Embraced herein, where applicable, are permissible isomers such astautomers, racemates, enantiomers, diastereomers, atropisomers,configurational isomers of double bonds (E- and/or Z-), cis- andtrans-configurations in ring substitution patterns, and isotopicvariants.

“Pharmaceutically acceptable” means suitable for use in pharmaceuticalpreparations, generally considered as safe for such use, officiallyapproved by a regulatory agency of a national or state government forsuch use, or being listed in the U.S. Pharmacopoeia or other generallyrecognized pharmacopoeia for use in animals, and more particularly inhumans.

“Pharmaceutically acceptable carrier” refers to a diluent, adjuvant,excipient, or carrier, or other ingredient which is pharmaceuticallyacceptable and with which a compound of the invention is administered.

“Pharmaceutically acceptable salt” refers to a salt which may enhancedesired pharmacological activity. Examples of pharmaceuticallyacceptable salts include acid addition salts formed with inorganic ororganic acids, metal salts and amine salts. Examples of acid additionsalts formed with inorganic acids include salts with hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid.Examples of acid addition salts formed with organic acids such as aceticacid, propionic acid, hexanoic acid, heptanoic acid,cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid,malonic acid, succinic acid, malic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, o-(4-hydroxy-benzoyl)-benzoicacid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonicacid, 1,2-ethanedisulfonic acid, 2-hydroxyethane-sulfonic acid,benzenesulfonic acid, p-chlorobenzenesulfonic acid,2-naphthalenesulfonic acid, p-toluenesulfonic acid, camphorsulfonicacid, 4-methyl-bicyclo[2.2.2]oct-2-enel-carboxylic acid, gluco-heptonicacid, 4,4′-methylenebis(3-hydroxy-2-naphthoic) acid, 3-phenylpropionicacid, trimethyl-acetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxy-naphthoic acids, salicylicacid, stearic acid and muconic acid. Examples of metal salts includesalts with sodium, potassium, calcium, magnesium, aluminum, iron, andzinc ions. Examples of amine salts include salts with ammonia andorganic nitrogenous bases strong enough to form salts with carboxylicacids.

A compound of a given formula (e.g. the compound of Formula I, whichalso includes Formula I, II, II-a, II-b, II-c and/or III) is intended toencompass the compounds of the disclosure, and the pharmaceuticallyacceptable salts, stereoisomers, mixture of stereoisomers or tautomersof such compounds. Additionally, the compounds of the disclosure maypossess one or more asymmetric centers, and can be produced as a racemicmixture or as individual enantiomers or diastereoisomers. The number ofstereoisomers present in any given compound of a given formula dependsupon the number of asymmetric centers present (there are 2^(n)stereoisomers possible where n is the number of asymmetric centers). Theindividual stereoisomers may be obtained by resolving a racemic ornon-racemic mixture of an intermediate at some appropriate stage of thesynthesis or by resolution of the compound by conventional means. Theindividual stereoisomers (including individual enantiomers anddiastereoisomers) as well as racemic and non-racemic mixtures ofstereoisomers are encompassed within the scope of the presentdisclosure, all of which are intended to be depicted by the structuresof this specification unless otherwise specifically indicated.

“Isomers” are different compounds that have the same molecular formula.Isomers include stereoisomers, enantiomers and diastereomers.

“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space. Stereoisomers include enantiomers and diastereomers.

“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A 1:1 mixture of a pair of enantiomers is a“racemic” mixture. The term “(±)” is used to designate a racemic mixturewhere appropriate.

“Diastereoisomers” are stereoisomers that have at least two asymmetricatoms, but which are not mirror-images of each other.

The absolute stereochemistry is specified according to the Cahn IngoldPrelog R S system. When the compound is a pure enantiomer thestereochemistry at each chiral carbon may be specified by either R or S.Resolved compounds whose absolute configuration is unknown aredesignated (+) or (−) depending on the direction (dextro- orlaevorotary) that they rotate the plane of polarized light at thewavelength of the sodium D line.

Some of the compounds exist as tautomeric isomers or “tautomers”.Tautomeric isomers are in equilibrium with one another. For example,amide containing compounds may exist in equilibrium with imidic acidtautomers. Regardless of which tautomer is shown, and regardless of thenature of the equilibrium among tautomers, the compounds are understoodby one of ordinary skill in the art to comprise both amide and imidicacid tautomers. Thus, the amide containing compounds are understood toinclude their imidic acid tautomers. Likewise, the imidic acidcontaining compounds are understood to include their amide tautomers.

“Therapeutically-effective amount” refers to an amount of a compoundthat, when administered to a subject for treating a disease, issufficient to effect treatment for the disease. “Therapeuticallyeffective amount” can vary depending on the compound, the disease andits severity, the age, the weight, etc. of the subject to be treated.

The term “treating”, and grammatical equivalents thereof, when used inthe context of treating a disease, means slowing or stopping theprogression of a disease, or ameliorating at least one symptom of adisease, more preferably ameliorating more than one symptom of adisease. For example, treatment of a hepatitis C virus infection caninclude reducing the HCV viral load in an HCV infected human being,and/or reducing the severity of jaundice present in an HCV infectedhuman being.

Any formula or structure given herein, including Formula I, II, II-a,II-b, II-c or III, is also intended to represent unlabeled forms as wellas isotopically labeled forms of the compounds. An “isotope” may havestructures depicted by the formulas given herein except that one or moreatoms are replaced by an atom having a selected atomic mass or massnumber. Examples of isotopes that can be incorporated into compounds ofthe disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine and chlorine, such as, but not limited to ²H(deuterium, D), ³H (tritium), ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, ³⁵S,³⁶Cl and ¹²⁵I. Various isotopically labeled compounds of the presentdisclosure, for example those into which radioactive isotopes such as³H, ¹³C and ¹⁴C are incorporated. Such isotopically labeled compoundsmay be useful in metabolic studies, reaction kinetic studies, detectionor imaging techniques, such as positron emission tomography (PET) orsingle-photon emission computed tomography (SPECT) including drug orsubstrate tissue distribution assays or in radioactive treatment ofpatients.

The disclosure also included compounds of Formula I, II, II-a, II-b,II-c or III in which from 1 to n hydrogens attached to a carbon atomis/are replaced by deuterium, in which n is the number of hydrogens inthe molecule. Such compounds exhibit increased resistance to metabolismand are thus useful for increasing the half life of any compound ofFormula I, II, II-a, II-b, II-c or III when administered to a mammal.See, for example, Foster, “Deuterium Isotope Effects in Studies of DrugMetabolism”, Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compoundsare synthesized by means well known in the art, for example by employingstarting materials in which one or more hydrogens have been replaced bydeuterium.

Deuterium labeled or substituted therapeutic compounds of the disclosuremay have improved DMPK (drug metabolism and pharmacokinetics)properties, relating to distribution, metabolism and excretion (ADME).Substitution with heavier isotopes such as deuterium may afford certaintherapeutic advantages resulting from greater metabolic stability, forexample increased in vivo half-life, reduced dosage requirements and/oran improvement in therapeutic index. An ¹⁸F labeled compound may beuseful for PET or SPECT studies. Isotopically labeled compounds of thisdisclosure and prodrugs thereof can generally be prepared by carryingout the procedures disclosed in the schemes or in the examples andpreparations described below by substituting a readily availableisotopically labeled reagent for a non-isotopically labeled reagent. Itis understood that deuterium in this context is regarded as asubstituent in the compound of Formula I, II, II-a, II-b, II-c or Ill.

The concentration of such a heavier isotope, specifically deuterium, maybe defined by an isotopic enrichment factor. In the compounds of thisdisclosure any atom not specifically designated as a particular isotopeis meant to represent any stable isotope of that atom. Unless otherwisestated, when a position is designated specifically as “H” or “hydrogen”,the position is understood to have hydrogen at its natural abundanceisotopic composition. Accordingly, in the compounds of this disclosureany atom specifically designated as a deuterium (D) is meant torepresent deuterium.

Compounds

The present application provides a compound represented by Formula I:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein L¹, A¹, A²,X¹, R¹, R², R³, R^(4a), R^(4b), R⁵ and R^(6a), R^(6b) are as definedabove.

In one embodiment, provided is a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

A is a bond, —O—, —S(O)_(n)—, —NH—, —N((C₁-C₄)alkyl)- or(C₁-C₂)alkylene;

A¹ is (C₁-C₅)alkylene, (C₂-C₅)alkenylene, (C₂-C₅)alkynylene, arylene,heteroarylene, cycloalkylene, heterocycloalkylene, aryl(C₁-C₂)alkylene,heteroaryl(C₁-C₂)alkylene, cycloalkyl(C₁-C₂)alkylene orheterocycloalkyl(C₁-C₂)alkylene, wherein a sp³ carbon atom of A¹ isoptionally replaced by —O—, —S(O)_(n)—, —NH— or —N((C₁-C₄)alkyl)-, andwherein a sp³ or sp² carbon atom of A¹ is optionally substituted withone or more groups selected from halo, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, aryl, heterocycloalkyl, cycloalkyl, aryl(C₁-C₄)alkyl,cycloalkyl(C₁-C₄)alkyl, heterocycloalkyl(C₁-C₄)alkyl,arylheterocycloalkyl(C₁-C₄)alkyl, —OR⁹, —SR⁹, —S(O)R⁹, —S(O)₂R⁹ and—N(R⁹)₂;

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)— cycloalkylene, arylene,cycloalkylene, (C₁-C₃)alkylene, (C₂-C₃)alkenylene or (C₂-C₃)alkynylene,wherein A² is optionally substituted with one or more substituentsselected from OR⁹, —SR⁹, —S(O)R⁹, —S(O)₂R⁹, —N(R⁹)₂, halo,halo(C₁-C₄)alkyl, halo(C₁-C₄)alkoxy, cyano and (C₁-C₈)alkyl;

L¹ is —O—C(O)—, —O—CH₂—, —NR¹⁰—C(O)—, —NR¹⁰—CH₂—, —NH—C(R¹⁰)₂— or—NH—S(O)₂—, wherein each R¹⁰ is independently H, (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, (C₁-C₄)alkenyl or cycloalkyl;

X¹ is bond, —O—, —NH—, —N((C₁-C₄)alkyl)- or heterocycloalkylene;

R¹ and R² are independently H, (C₁-C₄)alkyl, (C₂-C₄)alkenyl,(C₂-C₄)alkynyl, halo, cyano or —C(O)—(C₁-C₄)alkyl; or

R¹ and R², when taken together with the carbon to which they are bothattached, form —C(═O)—, —C(═S)— or —C(═N(C₁-C₄)alkyl)-;

R³ is H or (C₁-C₄)alkyl which is optionally substituted with halo,cyano, hydroxyl or (C₁-C₄)alkoxy;

R^(4a) and R^(4b) are independently H, (C₁-C₈)alkyl, aryl,aryl(C₁-C₄)alkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl,cycloalkyl or cycloalkyl(C₁-C₄)alkyl, wherein each of R^(4a) and R^(4b)is optionally substituted with one or more substituent selected fromcyano, —COOH, halo, hydroxyl, amino, mono(C₁-C₈)alkylamino,di(C₁-C₈)alkylamino, aryl and heteroaryl;

R^(5a) and R^(5b) are independently H, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, (C₁-C₈)alkoxy, aryl, heterocycloalkyl, cycloalkyl,aryl(C₁-C₄)alkyl, cycloalkyl(C₁-C₄)alkyl orheterocycloalkyl(C₁-C₄)alkyl, wherein R⁵ is optionally substituted withone or more substituent selected from —N₃, cyano, —COOH, halo, hydroxyl,amino, mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, aryl and heteroaryl,or

R^(5a) and R^(5b) together form a spirocycle having Formula (a):

wherein a carbon ring atom of Formula (a) is optionally substituted withone or more heteroatom selected from SO, SO₂, O and N, and wherein acarbon ring atom of Formula (a) optionally has one or more substituentsselected from halo, hydroxyl, —NH₂, (C₁-C₄)alkyl and (C₁-C₄)alkoxy;

R^(6a) and R^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituent selected from halo,hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, aryl, cycloalkyl,heterocycloalkyl, mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹,—NHC(O)R⁹ and (C₁-C₈)alkanoyl; or R^(6a) and R^(6b) together form aspirocycle having Formula (a);

each R⁸ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl, (C₂-C₄)alkynyl, aryl, heteroaryl, heterocycloalkyl orcycloalkyl, wherein R⁸ is optionally substituted with —OR, —N(R⁹)₂,—CON(R⁹)₂, or cyano;

each R⁹ is independently H, (C₁-C₄)alkyl, (C₂-C₄)alkenyl or(C₂-C₄)alkynyl;

each n is independently 0, 1 or 2; and

m is 1, 2, 3, 4 or 5.

In one aspect of the embodiment, A¹ is ethenylene, propenylene,butenylene, ethylene, propylene, butylene, oxypropylene, oxypropenylene,pyrazolylene, phenylene or pyrimidinylene.

In another aspect of the embodiment, A² is —CH(R⁸)-quinolinylene,—CH(R⁸)— isoquinolinylene, —CH(R⁸)-naphthyridinylene,—CH(R⁸)-cinnolinylene, —CH(R⁸)-quinoxalinylene, —CH(R⁸)-phenylene,—CH(R⁸)-naphthylene or —CH(R⁸)-halophenylene. In various aspects of theembodiment, A² is selected from the group consisting of

wherein the left bond of A⁵ linker is attached to X¹. In certainembodiments, R⁸ is methyl.

In some embodiments, X¹ is —O—, —NH— or —N(CH₃)—; R¹ and R², when takentogether with the carbon to which they are both attached, form —C(═O)—;and R³ is H. In another embodiments, X¹ is —O—, —NH— or —N(CH₃)—; R¹ andR², when taken together with the carbon to which they are both attached,form —C(═O)—; R³ is H; one of R^(4a) and R^(4b) is H and the other ismethyl; and one of R^(5a) and R^(5b) is H and the other is iso-propyl.

In one embodiment, X¹ is —O—, —NH— or —N(CH₃)—; R¹ and R², when takentogether with the carbon to which they are both attached, form —C(═O)—;R³ is H; one of R^(4a) and R^(4b) is H and the other is methyl; one ofR^(5a) and R^(5b) is H and the other is iso-propyl; A² is —CH(R⁸)—heteroarylene, and R⁸ is methyl.

In another embodiment, X¹ is —O— or —NH—; R¹ and R², when taken togetherwith the carbon to which they are both attached, form —C(═O)—; R³ is H;R^(4a) is H; R^(4b) is methyl; R⁵ is isopropyl; and R⁸ is methyl.

In one embodiment, one of R^(6a) and R^(6b) is H; and the other H,hydroxyl, —CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), (C₁-C₈)alkyl or (C₁-C₈)alkoxy,wherein R^(6a) or R^(6b) is optionally substituted with one or moresubstituents selected from the group consisting of halo, hydroxyl,(C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, heterocycloalkyl, (C₁-C₄)alkanoyl ordi(C₁-C₄)alkylamino. In another embodiment, R^(6a) is H; and R^(6b) isH, (C₁-C₈)alkyl or (C₁-C₈)alkyl substituted with heterocycloalkyl,(C₁-C₄)alkanoyl or di(C₁-C₄)alkylamino.

In one embodiment, L¹ is —O—C(O)—. In another embodiment, L¹ is —O—CH₂—.In yet another embodiment, L¹ is —N(CH₃)—C(O)—. In still anotherembodiment, L¹ is —NH—CH₂—. In certain embodiments, L¹ is —NH—S(O)₂—.

In another aspect of the embodiment, X¹ is 4- to 7-memberedheterocycloalkylene. In a particular embodiment, the heterocycloalkylenehas a N ring atom which is bonded to —C(R¹)(R²)—, and non-limitingexamples include:

In another embodiment, there is provided a compound of Formula II:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein:

A¹ is

A² is —CH(R⁸)-arylene, —CH(R)-heteroarylene, —CH(R)-heterocycloalkylene,—CH(R⁸)— cycloalkylene, arylene or cycloalkylene;

X¹ is a bond, —CH₂—, —O—, —NH—, —N(CH₃)—,

R^(4a) is H, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, hydroxyl(C₁-C₈)alkyl,(C₁-C₄)alkoxy(C₁-C₈)alkyl or (C₁-C₈)alkyl;

R⁵ is H, (C₁-C₄)alkyl, halo(C₁-C₈)alkyl, hydroxy(C₁-C₄)alkyl,(C₁-C₄)alkoxy(C₁-C₈)alkyl, cycloalkyl or heterocycloalkyl;

R^(6a) and R^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy, aryl, cycloalkyl, heterocycloalkyl,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹, —NHC(O)R⁹ and(C₁-C₈)alkanoyl where such an optional substitution is chemicallyfeasible; or R^(6a) and R^(6b) together form a spirocycle having Formula(a); or

R^(6a) and R^(6b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹;

R^(7a) and R^(7b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl;

R⁸ is H or (C₁-C₄)alky; and

each R⁹ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl or (C₂-C₄)alkynyl.

In certain embodiments of Formula II, A¹ is

wherein the left bond of A¹ linker is attached to A²;

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)— cycloalkylene, arylene orcycloalkylene, wherein the left bond of A² linker is attached to X¹;

X¹ is bond, —CH₂—, —O—, —NH—, —N(CH₃)—,

R^(4a) is H, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl;

R⁵ is H or (C₁-C₄)alkyl;

R^(6a) and R^(6b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl,

or

R^(6a) and R^(6b) together form

R^(7a) and R^(7b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl; and

R⁸ is H or (C₁-C₄)alkyl.

In certain embodiments of Formula II, R^(4a) is methyl; R⁵ isiso-propyl; R⁸ is methyl; and is

In certain embodiments of Formula II, R^(4a) is methyl; R⁵ isiso-propyl; R⁸ is methyl; and A² is

In one embodiment, R^(4a) is methyl; R⁵ is isopropyl; R⁸ is methyl; andA² is

Non-limiting examples of such compounds include the following compoundsand pharmaceutically acceptable salts thereof:

Formula (II-a)

Compound No. A¹ R^(6a) R^(6b) a-1 (Example 3) —CH═CH— H H a-2 (Example5) —CH═CH— Methyl H a-3 (Example 5) —CH═CH— H Methyl a-4 —CH═CH— MethylMethyl a-5 (Example 4) —CH═CHCH₂— Propyl H a-6 (Example 12) —CH═CHCH₂—Methyl Methyl a-7 (Example 13) —CH═CHCH₂—

H a-8 (Example 15) —CH═CHCH₂—

H a-9 (Example 16) —CH═CHCH₂—

H a-10 (Example 14) —CH═CHCH₂—

H a-11

Methyl H a-12 (Example 11) —CH═CHCH(CH₃)— —CH₃ H a-13 (Example 11)—CH═CHCH(CH₃)— H —CH₃ a-14 (Example 9) —CH₂CH₂CH₂— Propyl H wherein theleft bond of A¹ linker is attached to A².

A² linker of the compounds above can be replaced by various positionisomers. Non-limiting examples thereof include compounds having thefollowing formulae:

wherein the compounds of the formulae above have the same combination orpattern of substituents given in the table for Compounds a-1 to a-14.

Also included are compounds having the following formulae:

Formula (II-a2)

Compound No. A¹ R^(6a) R^(6b) a2-1 —CH═CH— H H a2-2 —CH═CH— Methyl Ha2-3 —CH═CH— H Methyl a2-4 —CH═CH— Methyl Methyl a2-5 —CH═CHCH₂— PropylH a2-6 —CH═CHCH₂— Methyl Methyl a2-7 —CH═CHCH₂—

H a2-8 —CH═CHCH₂—

H a2-9 —CH═CHCH₂—

H a2-10 —CH═CHCH₂—

H a2-11 (Example 2)

Methyl H a2-12 —CH═CHCH(CH₃)— —CH₃ H a2-13 —CH═CHCH(CH₃)— H —CH₃ a2-14—CH₂CH₂CH₂— Propyl H wherein the left bond of A¹ linker is attached toA².

A² linker of the compounds above can be replaced by various positionisomers. Non-limiting examples thereof include compounds having thefollowing formulae:

wherein the compounds of the formulae above have the same combination orpattern of substituents given in the table for Compounds a2-1 to a2-14.

In another aspect of the embodiment, R^(4a) is methyl; R⁵ is isopropyl;R⁸ is methyl; and A² is

Non-limiting examples of such compounds include the following compoundsand pharmaceutically acceptable salts thereof:

Formula (II-b)

Compound No. A¹ R^(6a) R^(6b) b-1 (Example 6)

H H b-2

Methyl H b-3

H H b-4 (Example 1)

Methyl H b-5 (Example 8) —CH═CH—(CH₃)₃— H H b-6 —CH═CH—(CH₃)₃— Methyl Hb-7

H H b-8 (Example 7)

Methyl H b-9

H H b-10 (Example 10)

Methyl H

In another embodiment, is provided a compound of Formula II-c:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein:

R^(6a) and R^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy, aryl, cycloalkyl, heterocycloalkyl,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹, —NHC(O)R⁹ and(C₁-C₈)alkanoyl where such an optional substitution is chemicallyfeasible; or R^(6a) and R^(6b) together form a spirocycle having Formula(a); or

R^(6a) and R^(6b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹;

each R⁹ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl or (C₂-C₄)alkynyl.

In one embodiment of the compounds described herein, R^(6a) and R^(6b)are independently H, —OH, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy,(C₁-C₈)alkyl, halo(C₁-C₈)alkyl, hydroxy(C₁-C₈)alkyl,(C₁-C₄)alkoxy(C₁-C₈)alkyl, halo(C₁-C₄)alkoxy(C₁-C₈)alkyl,

In one embodiment of the compounds described herein, R^(6a) and R^(6b)together from

In some embodiments, R^(6a) and R^(6b) together form

In one embodiment, the compound is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.

In another embodiment, the compound is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.

In yet another embodiment, the compound is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.

In one embodiment, the compound is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.

In certain embodiments, there is provided a compound of Formula III:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein

L¹ is —O—CH₂—, —N(CH₃)—C(O)—, —NH—CH₂—, —N(CH₃)—CH₂—, —NH—CH(CF₃)— or—NH—S(O)₂—;

A¹ is

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)— cycloalkylene, arylene orcycloalkylene;

X¹ is —O—, —N(CH₃)— or —NH—;

R^(4a) is H, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl;

R⁵ is H or (C₁-C₄)alkyl;

R^(6a) and R^(6b) are independently H, hydroxyl,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), (C₁-C₈)alkyl or (C₁-C₈)alkoxy, optionallysubstituted with one or more substituents selected from the groupconsisting of halo, hydroxyl, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy,heterocycloalkyl, (C₁-C₄)alkanoyl or di(C₁-C₄)alkylamino; or

R^(6a) and R^(6b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, halo, hydroxyl, —NH₂, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and —S(O)₂R⁹;

R^(7a) and R^(7b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl; and

R⁸ is H or (C₁-C₄)alkyl.

In some embodiments of Formula III, L¹ is —O—CH₂—, —N(CH₃)—C(O)—,—NH—CH₂— or —NH—S(O)₂—;

A¹ is

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)— cycloalkylene, arylene orcycloalkylene;

X¹ is —O—, —N(CH₃)— or —NH—;

R^(4a) is H, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl;

R⁵ is H or (C₁-C₄)alkyl;

R^(6a) and R^(6b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl,

R^(6a) and R^(6b) together form

R^(7a) and R^(7b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl; and

R⁸ is H or (C₁-C₄)alkyl.

In one embodiment, L¹ is —NH—CH₂—; R^(4a) is methyl; R⁵ is iso-propyl;R⁸ is methyl; and A²

In one embodiment, L¹ is —NH—S(O)₂—; R^(4a) is methyl; R⁵ is iso-propyl;R⁸ is methyl; and A²

In various aspects of this embodiment, A² is selected from

In one embodiment, L¹ is —N(CH₃)—C(O)—; R^(4a) is methyl; R⁵ isiso-propyl; R⁸ is methyl; and A² is

In one embodiment, L¹ is —O—CH₂—; R^(4a) is methyl; R⁵ is iso-propyl; R⁸is methyl; and A² is

In one aspect of the embodiment, L¹ is —NH—CH₂—; R^(4a) is methyl; R⁵ isisopropyl; and R⁸ is methyl. Non-limiting examples of such compoundsinclude the following compounds and pharmaceutically acceptable saltsthereof:

In one aspect of the embodiment, L¹ is —NH—S(O)₂—; R^(4a) is methyl; R⁵is isopropyl; and R⁸ is methyl. Non-limiting examples of such compoundsinclude the following compounds and pharmaceutically acceptable saltsthereof:

In one aspect of the embodiment, L¹ is —N(CH₃)—C(O)—; R^(4a) is methyl;R⁵ is isopropyl; and R⁸ is methyl. Non-limiting examples of suchcompounds include the following compounds and pharmaceuticallyacceptable salts thereof:

In one aspect of the embodiment, L¹ is —O—CH₂—; R^(4a) is methyl; R⁵ isisopropyl; and R⁸ is methyl. Non-limiting examples of such compoundsinclude the following compounds and pharmaceutically acceptable saltsthereof:

One skilled in the art will recognize that substituents and othermoieties of the compounds of the generic formulae herein should beselected in order to provide a compound which is sufficiently stable toprovide a pharmaceutically useful compound which can be formulated intoan acceptably stable pharmaceutical composition. Compounds which havesuch stability are contemplated as falling within the scope of thepresent invention. It should be understood by one skilled in the artthat any combination of the definitions and substituents described aboveshould not result in an inoperable species or compound.

Preparation of Macrocyclic Compounds

A compound of the present invention such as those of Formula I, II,II-a, II-b, II-c or III can be prepared according to the schemesdescribed below, but it shall be appreciated that modifications of theillustrated process or other process can also be used. As illustrated inScheme 1, the macrocyclic compounds M are synthesized from the five keycomponents A-E by combining them together in sequence with theappropriate use of protecting groups (PG₁-PG₈) by one skilled in theart. The hashed lines numbered 1-5, hereby referred to as Connection 1,Connection 2, etc, respectively, are the 5 connections for combiningComponents A-E. The order in which the specific connections occur, canvary, and are dependent on the choice of protecting groups and chemistryrequired. Typically Connections 3, 4 or 5 are used as the finalmacrocylization step.

Illustratively, Connections 1 through 5 can be performed as describedbelow:

Connections 1 and 2 are amide bonds. The connections are made betweenthe respective acid and amine using standard peptide coupling agents(EDC/HOBT, DCC, PyBOP, PyBROP, HATU, HBTU, COMU, etc) known to oneskilled in the art. The acid and amine coupling partners are combinedwith the coupling agent in an organic solvent, e.g., DMF,dichloromethane, acetonitrile, etc, in the presence of a base, e.g.,DIPEA, triethylamine, etc, at room temperature or slightly elevatedtemperature. When either of these steps is chosen as the finalmacrocyclization step, then macrolactamization conditions are preferred.Suitable macrolactamization procedures include, but are not limited to,those found in the following reference: Davies, J. S. J. Peptide Sci.2003, 9, 471-501.

Connection 4 is typically a carbon-carbon bond or a heteroatom-carbonbond where the heteroatom is O, S or N. When Connection 4 is acarbon-carbon bond, then standard carbon-carbon bond forming procedurestypically involving metal mediated cross coupling reactions arepreferred. Preferably the carbon-carbon bond is formed using a Heck typecoupling between an sp2 halide group and an terminal alkene, a Suzukicoupling between an sp2 halide group and a vinyl or aryl boronate, or aring closing metathesis (RCM) between two alkenes. Stille reactions canalso be performed between a vinyl stannane and an aryl or vinyl halideas described in Journal of American Chemical Society 2000, 122, 3830Nicolaou et al. In each of the examples above the aryl or vinyl halidegroup can also be an aryl or vinyl triflate.

For example, when Y in A is an alkene, preferably —CH═CH₂, and A₁ in Bcontains a terminal alkene or Me-CH═CH—, then a cross metathesisreaction is performed. The two components are mixed in solvent, e.g.,acetonitrile, toluene and a metathesis catalyst, e.g., Grubbs I, GrubbsII or Hoyveda-Grubbs I, Hoyveda-Grubbs II is added followed by heating.If this connection is the final procedure to close the macrocyclic ring,RCM conditions are preferred (e.g., more dilute conditions to avoiddimerization). For relevant RCM conditions and examples see Journal ofAmerican Chemical Society 2003, 125, 3849 Sedrani et al and Journal ofAmerican Chemical Society 2000, 122, 3830 Nicolaou et al. A typical RCMprocedure includes heating (either conventionally or by microwave) ofthe acyclic precursor in a solvent such as toluene, or1,2-dichloroethane, in the presence of a RCM catalyst, e.g., Grubbs I,Grubbs II or Hoyveda-Grubbs I, Hoyveda-Grubbs II.

Alternatively, when connection 4 is made via a Heck coupling reaction,the vinyl or aryl halide, or the triflate A and the alkene component Bare mixed in a polar solvent, e.g., acetonitrile or toluene, in thepresence of a Palladium(II) catalyst, e.g., Palladium(OAc)₂, a phosphineligand, e.g., P(o-toluene)₃, P(t-butyl)₃, etc, and a base, e.g.,triethylamine. The reaction mixture is heated either conventionally orin a microwave reactor.

Alternatively, when Connection 4 is made via a Suzuki coupling reaction,the vinyl or aryl halide, or the triflate A and the vinyl or arylboronate B are mixed in a suitable solvent, e.g., cyclopentyl methylether, toluene, DMF, DME, etc., in the presence of a Palladium catalyst(e.g., Palladium(II)Cl₂(p-NMe₂Ph)₂ and K₃PO₄ ortetrakis(triphenylphosphine)palladium(0) and a base, such as potassiumcarbonate). The reaction mixture is heated either conventionally or in amicrowave reactor. It is also possible in such a coupling reaction toreverse the reactive functionalities on the two starting materials, suchthat A is an aryl or vinyl boronate and B contains a vinyl or arylhalide or triflate.

Alternatively, Connection 4 can be a carbon-oxygen bond and in thiscase, typical alkylation or nucleophilic aromatic substitutionconditions can be used between a hydroxyl group and an alkyl halide, oraryl (or heteroaryl) halide. The hydroxyl reagent is mixed with thealkyl or heteroaryl halide (preferably an iodide or bromide), in aninert solvent, e.g., CPME, DMF, THF, etc, in the presence base, e.g.,cesium carbonate, cesium hydroxide, sodium hydride, NaHMDS, etc, andheated.

Alternatively, Connection 4 can be a carbon-nitrogen bond and in thiscase, typical alkylation, nucleophilic aromatic substitution or Buchwaldconditions can be used between an amine group and an alkyl halide orheteroaryl halide. For example, the amine and the alkyl or heteroarylhalide are mixed and heated in an inert solvent, e.g., CPME, in thepresence base, e.g., cesium carbonate, sodium hydride etc. Analternative procedure for the carbon-nitrogen connection is to perform areductive amination between an amine and a carbonyl compound. Typicallythe amine and aldehyde or ketone are mixed in an inert solvent, e.g.,THF, dioxane and treated after a period of time with sodium acetoxyborohydride or alternative reducing agent.

Connection 5 is typically an amide (X₁ is NH or substituted N), or ester(X₁ is O) bond. When forming the amide bond, standard couplingprocedures described for Connection 1-3 can be used. Often, this is thefinal step in closing the macrocycle. As such, macrolactamization typesof coupling procedures are more effective. Suitable macrolactamizationprocedures include, but are not limited to, those found in the followingreference: Davies, J. S. J. Peptide Sci. 2003, 9, 471-501.

When forming the ester bond coupling reagents (e.g., EDC, DCC, PyBOP,HATU, COMU) can be used, or when this is the final step in formation ofthe macrocycle, macrolactonization procedures are preferred (e.g.,Shiina, Yamaguchi). An example method for the macrolactonization stepcan be found in Journal of American Chemical Society 2002, 124, 4257Paquette et al or Chemical Reviews 2006, 106(3), 911-939. Typically, theacid and alcohol are mixed in a polar solvent, e.g., DMF, acetonitrile,etc. in the presence of the coupling agent and a base, e.g., DIPEA,DMAP.

Connection 5 can also be an ether bond where R¹ and R² are both hydrogenor alkyl groups. In this example, the carbon-oxygen bond formingprocedures described above for Connection 4 are used to connect the 2components. Connection 5 can also be an amine bond where R¹ and R² areboth hydrogen or alkyl groups and the procedures for connecting theamine and carbonyl components are also described above with respect toConnection 4.

Connection 3 is a C—N, C—O, or S(O)₂—N. Suitable bond forming reactionsfor Component C and Component B include the following methods.

When the bond formed in Connection 3 is L¹ is —OC(O)— (i.e., X¹ is O inC and Z/Z¹ is —CO₂H or —CO-halogen in B), this is performed usingstandard ester formation procedures utilizing coupling reagentsdescribed above. When this is the final step in formation of themacrocycle, macrolactonization procedures are preferred, as describedabove.

Alternatively, when Connection 3 is L¹ is —N(alkyl)C(O)— (i.e., X¹ isN(alkyl) in C and Z/Z¹ is CO₂H or CO-halogen in B) this step isperformed using standard ester formation procedures utilizing couplingreagents described above. When this is the final step in formation ofthe macrocycle, macrolactamization procedures are preferred as describedabove.

Alternatively, when connection 3 is L¹ is —NHS(O)₂— (i.e., X¹ is —NH— inC and Z/Z¹ is —S(O)₂OH or —S(O)₂-halogen in B), this is performed usingstandard amide formation procedures utilizing coupling reagentsdescribed above, or when this is the final step in formation of themacrocycle, macrolactamization procedures are preferred as describedabove.

Alternatively, when Connection 3 is L¹ is —NH—CH₂— (i.e., X¹ is —NH— inC and Z/Z¹ is ketone or aldehyde in B), this is performed using standardreductive amination procedures as described above.

Alternatively, when Connection 3 is L¹ is —O—CH₂— (i.e., X¹ is O in Cand Z/Z¹ contains a leaving group such as a halide or triflate in B),this is performed using standard nucleophilic alkylation proceduresdescribed above.

The following general schemes provide general examples and sequences forconstructing the macrocyclic compounds M from the common precursors A-E.

Compounds A-E are first deprotected (PG²-PG⁸) using conditions describedin Greene and Wuts, Protective Groups in Organic Synthesis, John Wileyand Sons, Inc. to provide Compounds 1a-1f.

In many cases the optimal protecting groups and their deprotectionmethods are as follows. For Compound E the typical protecting group PG¹for the acid (when R¹ and R² are C═O) is a methyl or trichloroethylester. The methyl and trichloroethyl esters can be removed by base,e.g., LiOH in a polar solvent, e.g., aqueous THF, etc. Thetrichloroethyl ester can also be removed by treatment with zinc andammonium acetate in a polar solvent, e.g., THF. Typically, PG² and PG⁴are acid labile groups, e.g., BOC and are deprotected using HCl indioxane or TMSOTf in dioxane, dichloromethane. Typically, PG³ and PG⁵are ester groups, removed by treatment with alkali metal hydroxide inaqueous THF or dioxane. Typically PG⁶ is an acid labile group, e.g., BOCfor amine and removed as described for PG²; or silyl ether for ahydroxyl group and removed by treatment with HF. pyridine or TBAF in anorganic solvent, e.g., dichloromethane. Typically PG⁸ is an amineprotecting group, e.g., BOC and removed as described for PG² or a silylether of a hydroxyl group removed as described for PG⁶, or an acetateprotecting group removed by treatment with alkali metal hydroxide inaqueous THF or dioxane.

Compound 1f is coupled to acid 1a using the conditions described abovefor connection 1 to produce Compound 1g. Compound 1g is then PG⁴deprotected using conditions described in Greene and Wuts and coupled toacid 1c to provide 1 h using the conditions described above forConnection 2. An alternative sequence for generating 1 h, is firstcoupling amine 1b to acid 1c using conditions as described forconnection 2 above, to form ii; deprotection of the protecting group PG³in ii using conditions described in Greene and Wuts, and finallycoupling with amine 1f using conditions described for Connection 1 aboveto form 1h.

Compound 1h is deprotected at PG⁶ using conditions described in Greeneand Wuts and is then coupled to B using the conditions described abovefor connection 3 to form 1j. An alternative route to 1j is alsoillustrated. Component B is coupled to 1d using the methods describedabove for Connection 3. The protecting group PG⁵ is then removed usingconditions described in Greene and Wuts and the product coupled with thedipeptide produced from PG⁴ deprotected Ig, to provide 1j. Protectinggroup PG¹ in Compound 1j is then removed using conditions described inGreene and Wuts, and the acid is then coupled to 1e using conditionsdescribed for Connection 5 to form the intermediate 1k. An alternativesequence to 1k from the intermediate 1 h is first, deprotection of PG¹and then coupling to 1e as described for Connection 5; and thendeprotection of PG⁶ as described in Greene and Wuts, followed bycoupling to B using the conditions described for Connection 3 to form1k. Product 1k is then subjected to the RCM as described above forConnection 4 to form Compound M.

Compound A is coupled to Compound B using the conditions described abovefor Connection 4 to generate 2a. Typically in this sequence Compound Bwould be protected, e.g., an acid would be an ester, a sulfonic acid, asulfonate ester, a ketone or aldehyde an acetal for example. Compound 2ais then deprotected as described in Greene and Wuts to generate acid 2b.Acid 2b is then coupled to the deprotected product of 1 h (prepared from1h by deprotection of PG⁶ described in Greene and Wuts) to generate theprecursor 2c. Deprotection of 2c at both PG⁸ and PG¹ is carried outusing conditions described in Greene and Wuts, and then the product iscyclized using the conditions described above for macrolactamization ormacrolactonization in Connection 5, to provide Compound M.

Compound 2a is deprotected at PG₈ as described in Greene and Wuts togenerate 3a which is then coupled, using conditions described above forconnection 5, to the PG¹ deprotected product of 1h, (prepared from 1h bydeprotection of PG¹ described in Greene and Wuts), to generate theprecursor 3b. Deprotection of 3b at both PG⁶ and Z/Z¹ if required iscarried out using conditions described in Greene and Wuts, and then theproduct is cyclized using the conditions described above formacrolactamization or macrolactonization in Connection 3, to provideCompound M.

The final macrocycle M from Schemes 4-6 often contains functionalitiesof protecting groups on sidechains that require further removal togenerate the final compound M. For example, when M contains a C═C as aresult of RCM Compound M is mixed in a solvent such as ethanol,methanol, etc, in the presence of palladium on carbon catalyst under anatmosphere of hydrogen gas to provide reduced Compound M1. Protectinggroups on the R^(4a), R^(4b), R⁵, A¹, A² and/or X¹ are removed usingconditions described in Greene and Wuts to generate Compound M2. Anothertransformation is click chemistry, to produce triazole M3. Thistransformation is performed by treating the alkyne or azide in M, insolvent (e.g., DMF) with an alkyne or azide as appropriate in thepresence of CuI to form M3.

Deprotected compound M2 can be further transformed after deprotection toadditional macrocycle M, for example: Treatment of M2 containing OH withan alkyl halide in the presence of base, e.g., cesium carbonate, insolvent, e.g., DMF, acetonitrile forms alkylated product M4. M2containing a ketone group is treated in solvent (e.g., DMF, methanol,etc) with an amine followed by the addition of sodium acetoxyborohydrideto form amine product M5.

Many acids, esters, ketones, aldehyde, sulfonic acid or halide componentB with a terminal alkene, Me-CH═C—, or vinyl/aryl boronate, or halide inA1 are commercially available or described in the literature and thesecan be used in the above schemes directly. In addition, the followingschemes below are examples of methods that can be used by one skilled inthe art to generate additional B.

In Scheme 8, part 1, a protected acid is treated with a strongdeprotonating base, e.g., LDA in an inert solvent, e.g., THF at −78° C.and HMPA. A pre-cooled solution of dichloroacteylene (prepared bytreatment of trichlorethene with potassium hydride and MeOH (catalytic)in THF) is then added to generate the chloro acetylene product. Thisproduct is then reduced, for example, by treatment with Cu in aceticacid and THF to generate the alkyne which is then further reduced to thealkene, for example, by treatment of an alcoholic solution of the alkynewith a poisoned palladium reducing agent (e.g. Lindlar) in the presenceof hydrogen gas. Alternatively the alkyne is treated with Cp₂ZrHCl indichloromethane, in the presence of pinnacolborane to form the vinylboronate.

In Scheme 8, part 2 a beta-keto ester with alpha substitution isconverted to the vinyl triflate, for example by treating a THF solutionof the beta-keto ester with a base, e.g., LDA in THF at −78° C.,followed by addition of PhN(Tf)₂. The triflate product is then treatedwith pyridine at elevated temperature to form the alkyne. The alkyne isthen treated as described above in Example 1 to generate the alkene orvinyl boronate products.

In Scheme 8, part 3, a chiral aldol reaction is used. An acyl group isfirst attached to a chiral auxiliary, e.g., Evans, Oppolzer sultam (seeJACS 1990, 112, p 2767) using standard amide bond formation conditionsdescribed for connection 1-3. The Oppolzer auxiliary product is treatedwith the aldehyde of choice, TBDMSOTf and base, e.g., triethylamine inanhydrous solvent, e.g., dichloromethane. The Evans auxiliary is treatedwith base, e.g., LDA, KHMDS, DIPEA in organic solvent, e.g., THF at −78°C. and the aldehyde of choice in the presence of a Lewis acid, e.g.,TiCl₄, SnCl₄ or BF₃OEt₂. Protection of the resulting alcohol from thealdol reaction is performed as described in Greene and Wuts, oralternatively alkylation with an alkyl halide or Meerwein's reagent,i.e. treatment with trimethyloxonium tetrafluoroborate in an inertsolvent, e.g., dichloromethane is performed. The auxiliary is thenremoved using standard alkali metal hydroxide removal conditions, e.g.,LiOH in THF, or LiOH and hydrogen peroxide in THF to provide the freeacids product.

In Scheme 8, part 4, an Evans auxiliary is allylated with an allylhalide as described in Synlett 2002, 12, 2039-2040. The product is thenisomerized by treatment with RhCl₃ in ethanol and then the auxiliaryremoved by base and peroxide, e.g., LiOH and H₂O₂ in THF/Water.Alternatively the auxiliary is directly removed by LiOH and H₂O₂ inTHF/Water to provide the terminal alkene.

In Scheme 8, part 5, a Horner Wadsworth Emmons reaction is used on analdehyde (containing a terminal alkene) to generate the alpha-betaunsaturated ester which is then selectively reduced to the ester. Forexample the phosphonate is treated with base, e.g., sodium hydride inTHF at low temperature, followed by addition of the aldehyde and warmingto generate the unsaturated ester. The product is reduced by treatmentwith magnesium powder in methanol.

In Scheme 8, part 6, an alpha-beta unsaturated acid or is converted tothe unsaturated Evans auxiliary (see Organic Letters 2007, 9, p 1635)and is treated with an aldehyde to generate the corresponding alkeneproduct. The hydroxyl group is then protected using methods described inGreene and Wuts and then the auxiliary is removed by treatment with baseand peroxide, e.g., LiOH and H₂O₂ in THF/Water. The hydroxyl can also bealkylated by as described above for aldol Scheme 8, part 3.

In Scheme 8, part 7 a ketone is transformed via the nitro olefin asdescribed in Angew. Chem. Int. Ed. 2006, 45 (46), 7736. The nitro olefinis then treated with vinyl magnesium bromide in an inert solvent, e.g.,THF, in the presence of a copper(I)salt, e.g., CuI and trimethylsilylchloride. The nitro alkyl product after addition of the vinyl group isthen converted to the acid by treating with sodium nitrite and aceticacid in an inert polar solvent, e.g., DMSO.

To generate the ketones, aldehydes, hydroxy, or halide compounds theacid/ester products described in Scheme 8 can be transformed usingstandard methods known to one skilled in the art. For example,conversion of the acid to the Weinreb amide with a coupling agent in asolvent, e.g., THF, dichloromethane, followed by treatment with anucleophile, e.g., MeMgBr, CF₃SiMe and the like, generates ketones;hydroxyl groups are produced by treatment of the esters with lithiumaluminum hydride in a solvent, e.g., THF, dichloromethane etc. Hydroxylsare activated to triflates with treatment with triflic anhydride, and abase, e.g., diisopropylethylamine in solvent, e.g., THF. Conversion ofthe hydroxyl to a halide, e.g., bromide is performed by treatment withPPh₃ and carbon tetrabromide.

Several types of A (or A1 as shown) are available commercially ordescribed in the literature where X¹ is O or NH and Y is a halide oralkene. The schemes below describe additional general methods forgenerating A1.

In Scheme 9, part 1 (Y is a halogen in A1); the starting compound 6a istypically a commercially available aromatic compound, that containshalogen Y and a group Z that can be transformed to the ketone 6b.Typical Z groups are halide, acid, aldehyde, for example.

When Z is an acid, 6a is treated with a coupling agent, e.g., HATU, EDCin the presence of a base, e.g., DIPEA and the Weinreb amine (Me—NH—OMe)to form the Weinreb amide. The amide is then treated with a nucleophile,e.g., TMS-CF₃ to form the CF₃ substituted ketone 6b or with a Grignardagent, e.g., MeMgBr in a solvent, e.g., THF at −78° C. to form themethyl ketone 6b.

When Z is halogen, then the initial conversion, if required, to a morereactive halogen is performed by treatment with NaI and acetyl chloridein an inert solvent, e.g., acetonitrile. The halogen is then transformedto the ketone by a Stille reaction with an ethoxyvinyl stannane. Thehalide is treated in an inert solvent, e.g., toluene, with the stannaneand a palladium (II) catalyst, e.g., PdCl₂(PPh₃)₂, followed by treatmentof the product with 2M HCl to afford ketone 6b. In some cases, theformation of an alkyl lithium reagent from the halide group can beperformed by treatment with nBuLi at −78° C. in THF and then addition ofa N-methoxy-N-methyl amide to afford the ketone 6b (e.g.,N-methoxy-N-methylacetamide affords the R⁸=methyl ketone 6b). A finalmethod to generate the ketone 6b is through a vinyl group. 6a is treatedwith a vinyltrifluoroborate in the presence of a palladium catalyst,e.g., PdCl₂(dppf) and then the vinyl product is subsequently ozonolysedin a polar solvent, e.g., methanol at low temperature to give analdehyde. The aldehyde is then reacted with a nucleophile, e.g., TSM-CF₃or a Grignard reagent, e.g., MeMgBr to afford a secondary alcoholproduct. The secondary alcohol is then oxidized with Dess MartinPeriodinane to give the desired ketone 6b or can be used as A1 itself.

Chiral alcohol (X¹ is O) and amine (X¹ is NH) A1 are generated usingChiral reduction methods on the ketone 6b. Chiral alcohol 6c is formedfrom 6b using one of the numerous chiral reduction methods available inthe literature. Typically, dichloro(p-cumene)ruthenium(II) dimer and(1R,2R)-(−)—N-p-tosyl-1,2-diphenylethylenediamine are combined in water,and sodium formate and 6b is added in a water miscible solvent such astetrahydrofuran. The reaction is then stirred at a temperature betweenambient and reflux to produce 6c where X¹ is O. Alternatively, a chiralCBS reduction can be performed in an inert solvent, e.g., THF at lowtemperature to also afford the chiral alcohol 6c. Protection of the OHin 6c is performed using methods described in Greene and Wuts, typicallya TBS ether or acetyl group are used to provide A1 (X¹ is O).

Alternatively to make chiral A1 (X¹ is NH), ketone 6b is first convertedto a chiral imine (R¹⁰ is chiral group) and then reduced using a varietyof methods described in the literature. For example, a chiralsulfinamide is reacted with the ketone 6b to afford a chiral sulfinimine6d, which is then reduced with a suitable reducing agent, typicallyNaBH₄, or selectride, or a Noyori type reduction as described for thechiral alcohol above, with dichloro(p-cumene)ruthenium(II) dimer and(1R,2R)-(−)—N-p-tosyl-1,2-diphenylethylenediamine. The sulfinamideauxiliary is then removed by treatment with mineral acid, preferably HClin a suitable organic solvent such as methanol, to afford 6e whereX₁═NH. Protection of the NH group can then be performed as described inGreene and Wuts to generate A1 (X¹ is NH).

In part 2 of Scheme 9, the synthesis of compound A1 where Y is —CH═CH₂,a precursor for metathesis and cross coupling reactions is illustrated.Examplary methods are as follows.

Compound 6c or 6e generated in Scheme 9, part 1 is first optionallyprotected on X¹ using a suitable protecting group as described in Greeneand Wuts, and then a vinyl group is introduced by a suitable crosscoupling method onto the aryl or sp2 halide. For example, a transitionmetal mediated coupling with an vinyl stannane or vinyltetrafluoroborate using a suitable palladium catalyst, e.g.,PdCl₂(dppf)₂ or Pd Cl₂(PPh₃)₂ in a suitable organic solvent, e.g.,acetonitrile, dichloromethane, etc., with either thermal or microwaveheating affords alkene A1.

Another typical method that can be used to introduce a vinyl group isstarting from the ketone 6b where Y is OH. Initially, triflation of thealcohol is performed by treatment with Tf₂O in the presence of a base,e.g., pyridine. The ketone group is then reduced with a Noyorireduction, or as described above through the sulfonamide, to provide thechiral alcohol or amine. The chiral alcohol or amine is then protectedas described in Greene and Wuts, and then the triflate is reacted with avinyl cross coupling reagent, e.g., vinyl stannane in a Stille coupling,or a vinyltrifluoroborate as described above to introduce the alkene. Afurther example of alkene generation using 6b ketone is via introductionof an allyl group. Thus, 6b where Y is OH is treated in an inert solventin the presence of a suitable base, e.g., alkali metal carbonate,preferably potassium carbonate with allyl bromide to form 6f. Compound6f is then similar to ketone 6b and is therefore able to be transformedas described above in part 1 to compound A1 where X¹ is O or NH withprotecting group PG⁸.

Pharmaceutical Formulations

The compounds of this invention are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations of the invention, both for veterinary and for human use,comprise at least one active ingredient, together with one or moreacceptable carriers and optionally other therapeutic ingredients.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or a suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

The effective dose of an active ingredient depends at least on thenature of the condition being treated, toxicity, whether the compound isbeing used prophylactically (lower doses) or against an active viralinfection, the method of delivery, and the pharmaceutical formulation,and will be determined by the clinician using conventional doseescalation studies. The effective dose can be expected to be from about0.0001 to about 100 mg/kg body weight per day; typically, from about0.01 to about 10 mg/kg body weight per day; more typically, from about0.01 to about 5 mg/kg body weight per day; most typically, from about0.05 to about 0.5 mg/kg body weight per day. For example, the dailycandidate dose for an adult human of approximately 70 kg body weightwill range from 1 mg to 1000 mg, preferably between 5 mg and 500 mg, andmay take the form of single or multiple doses.

Combination Therapy

The compounds of the present invention may be combined with one or moreactive agents. Non-limiting examples of suitable active agents to becombined include one or more interferons, ribavirin or its analogs, HCVNS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase 1inhibitors, hepatoprotectants, mevalonate decarboxylase antagonists,antagonists of the renin-angiotensin system, other anti-fibrotic agents,endothelin antagonists, nucleoside or nucleotide inhibitors of HCV NS5Bpolymerase, non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5Ainhibitors, TLR-7 agonists, cyclophillin inhibitors, HCV IRESinhibitors, pharmacokinetic enhancers and other drugs for treating HCV;or mixtures thereof.

More specifically, one or more compounds to be combined are selectedfrom the group consisting of:

1) interferons, e.g., pegylated rlFN-alpha 2b (PEG-Intron), pegylatedrlFN-alpha 2a (Pegasys), rlFN-alpha 2b (Intron A), rlFN-alpha 2a(Roferon-A), interferon alpha (MOR-22, OPC-18, Alfaferone, Alfanative,Multiferon, subalin), interferon alfacon-1 (Infergen), interferonalpha-n1 (Wellferon), interferon alpha-n3 (Alferon), interferon-beta(Avonex, DL-8234), interferon-omega (omega DUROS, Biomed 510),albinterferon alpha-2b (Albuferon), IFN alpha XL, BLX-883 (Locteron),DA-3021, glycosylated interferon alpha-2b (AVI-005), PEG-Infergen,PEGylated interferon lambda (PEGylated IL-29), and belerofon;

2) ribavirin and its analogs, e.g., ribavirin (Rebetol, Copegus), andtaribavirin (Viramidine);

3) HCV NS3 protease inhibitors, e.g., boceprevir (SCH-503034, SCH-7),telaprevir (VX-950), VX-813, TMC-435 (TMC435350), ABT-450, BI-201335,BI-1230, MK-7009, SCH-900518, VBY-376, VX-500, GS-9256, GS-9451,BMS-790052, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530, YH5531, andITMN-191 (R-7227);

4) alpha-glucosidase 1 inhibitors, e.g., celgosivir (MX-3253), Miglitol,and UT-231B;

5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738, GS-9450(LB-84451), silibilin, and MitoQ;

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, e.g.,R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851, BCX-4678,valopicitabine (NM-283), GS-6620 and MK-0608;

7) non-nucleoside inhibitors of HCV NS5B polymerase, e.g., filibuvir(PF-868554), ABT-333, ABT-072, BI-207127, VCH-759, VCH-916, JTK-652,MK-3281, VBY-708, VCH-222, A848837, ANA-598, GL60667, GL59728, A-63890,A-48773, A-48547, BC-2329, VCH-796 (nesbuvir), GSK625433, BILN-1941,XTL-2125, and GS-9190;

8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), AZD-7295 (A-689), andBMS-790052;

9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773, ANA-975,AZD-8848 (DSP-3025), PF-04878691, and SM-360320;

10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and NIM811;

11) HCV IRES inhibitors, e.g., MCI-067;

12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452, PF-4194477,TMC-41629, GS-9350, GS-9585, and roxythromycin;

13) other drugs for treating HCV, e.g., thymosin alpha 1 (Zadaxin),nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir, GI-5005,XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOc-205, tarvacin,EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032, BMS-791325,Bavituximab, MDX-1106 (ONO-4538), Oglufanide, FK-788, and VX-497(merimepodib);

14) mevalonate decarboxylase antagonists, e.g., statins, HMGCoA synthaseinhibitors (e.g., hymeglusin), squalene synthesis inhibitors (e.g.,zaragozic acid);

15) angiotensin II receptor antagonists, e.g., losartan, irbesartan,olmesartan, candesartan, valsartan, telmisartan, eprosartan;

16) angiotensin-converting enzyme inhibitors, e.g., captopril,zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril,benazepril, fosinopril;

17) other anti-fibrotic agents, e.g., amiloride; and

18) endothelin antagonists, e.g. bosentan and ambrisentan.

In yet another embodiment, the present application provides acombination therapy comprising a composition of the present inventionand a second pharmaceutical composition comprising at least oneadditional therapeutic agent selected from the group consisting of HIVprotease inhibiting compounds, HIV non-nucleoside inhibitors of reversetranscriptase, HIV nucleoside inhibitors of reverse transcriptase, HIVnucleotide inhibitors of reverse transcriptase, HIV integraseinhibitors, gp41 inhibitors, CXCR4 inhibitors, gp120 inhibitors, CCR5inhibitors, interferons, ribavirin analogs, NS3 protease inhibitors,NS5a inhibitors, alpha-glucosidase 1 inhibitors, cyclophilin inhibitors,hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs fortreating HCV, and combinations thereof.

More specifically, one or more compounds of the present invention may becombined with one or more compounds selected from the group consistingof 1) HIV protease inhibitors, e.g., amprenavir, atazanavir,fosamprenavir, indinavir, lopinavir, ritonavir, lopinavir+ritonavir,nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126,TMC-114, mozenavir (DMP-450), JE-2147 (AG1776), AG1859, DG35, L-756423,R00334649, KNI-272, DPC-681, DPC-684, and GW640385X, DG17, PPL-100, 2) aHIV non-nucleoside inhibitor of reverse transcriptase, e.g.,capravirine, emivirine, delaviridine, efavirenz, nevirapine, (+)calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963, Mb-150, andTMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355 BS, VRX 840773,UK-453,061, RDEA806, 3) a HIV nucleoside inhibitor of reversetranscriptase, e.g., zidovudine, emtricitabine, didanosine, stavudine,zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine, alovudine,Mb-210, racivir (±-FTC), D-d4FC, emtricitabine, phosphazide, fozivudinetidoxil, fosalvudine tidoxil, apricitibine (AVX754), amdoxovir, KP-1461,abacavir+lamivudine, abacavir+lamivudine+zidovudine,zidovudine+lamivudine, 4) a HIV nucleotide inhibitor of reversetranscriptase, e.g., tenofovir, tenofovir disoproxilfumarate+emtricitabine, tenofovir disoproxilfumarate+emtricitabine+efavirenz, and adefovir, 5) a HIV integraseinhibitor, e.g., curcumin, derivatives of curcumin, chicoric acid,derivatives of chicoric acid, 3,5-dicaffeoylquinic acid, derivatives of3,5-dicaffeoylquinic acid, aurintricarboxylic acid, derivatives ofaurintricarboxylic acid, caffeic acid phenethyl ester, derivatives ofcaffeic acid phenethyl ester, tyrphostin, derivatives of tyrphostin,quercetin, derivatives of quercetin, S-1360, zintevir (AR-177),L-870812, and L-870810, MK-0518 (raltegravir), BMS-707035, MK-2048,BA-011, BMS-538158, GSK364735C, 6) a gp41 inhibitor, e.g., enfuvirtide,sifuvirtide, FB006M, TRI-1144, SPC3, DES6, Locus gp41, CovX, and REP 9,7) a CXCR4 inhibitor, e.g., AMD-070, 8) an entry inhibitor, e.g., SP01A,TNX-355, 9) a gp120 inhibitor, e.g., BMS-488043 and BlockAide/CR, 10) aG6PD and NADH-oxidase inhibitor, e.g., immunitin, 10) a CCR5 inhibitor,e.g., aplaviroc, vicriviroc, INCB9471, PRO-140, INCB15050, PF-232798,CCR5 mAb004, and maraviroc, 11) an interferon, e.g., pegylatedrlFN-alpha 2b, pegylated rlFN-alpha 2a, rlFN-alpha 2b, IFN alpha-2b XL,rlFN-alpha 2a, consensus IFN alpha, infergen, rebif, locteron, AVI-005,PEG-infergen, pegylated IFN-beta, oral interferon alpha, feron,reaferon, intermax alpha, r-IFN-beta, infergen+actimmune, IFN-omega withDUROS, and albuferon, 12) ribavirin analogs, e.g., rebetol, copegus,VX-497, and viramidine (taribavirin) 13) NS5a inhibitors, e.g., A-831,A-689 and BMS-790052, 14) NS5b polymerase inhibitors, e.g., NM-283,valopicitabine, R1626, PSI-6130 (R1656), IDX184, PSI-7851, HCc-796, BILB1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433, andXTL-2125, 15) NS3 protease inhibitors, e.g., SCH-503034 (SCH-7), VX-950(Telaprevir), ITMN-191, and BILN-2065, 16) alpha-glucosidase 1inhibitors, e.g., MX-3253 (celgosivir) and UT-231B, 17)hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and LB-84451, 18)non-nucleoside inhibitors of HCV, e.g., benzimidazole derivatives,benzo-1,2,4-thiadiazine derivatives, and phenylalanine derivatives, 19)other drugs for treating HCV, e.g., zadaxin, nitazoxanide (alinea),BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702, AVI 4065,bavituximab, oglufanide, PYN-17, KPE02003002, actilon (CPG-10101),KRN-7000, civacir, GI-5005, ANA-975, XTL-6865, ANA 971, NOc-205,tarvacin, EHC-18, and NIM811, 19) pharmacokinetic enhancers, e.g.,BAS-100, SPI-452, PF-4194477, TMC-41629, GS-9350, GS-9585, androxythromycin, 20)RNAse H inhibitors, e.g., ODN-93 and ODN-112, 21)other anti-HIV agents, e.g., VGc-1, PA-457 (bevirimat), ampligen,HRG214, cytolin, polymun, VGX-410, KD247, AMZ 0026, CYT 99007, A-221HIV, BAY 50-4798, MDX010 (iplimumab), PBS119, ALG889, and PA-1050040.

In a specific aspect of this embodiment, the additional therapeuticagent is selected from ribavirin, telaprevir, boceprevir and sofosbuvir(GS-7977 (formerly PSI-7977)).

A combination therapy described herein may be administered as asimultaneous or sequential regimen. When administered sequentially, thecombination may be administered in two or more administrations.

Co-administration of a compound of the invention with one or more otheractive agents generally refers to simultaneous or sequentialadministration of a compound of the invention and one or more otheractive agents, such that therapeutically effective amounts of thecompound of the invention and one or more other active agents are bothpresent in the body of the patient.

One or more compounds of the disclosure are administered by any routeappropriate to the condition to be treated. Suitable routes includeoral, rectal, nasal, topical (including buccal and sublingual), vaginaland parenteral (including subcutaneous, intramuscular, intravenous,intradermal, intrathecal and epidural), and the like. It will beappreciated that the preferred route may vary with for example thecondition of the recipient. An advantage of the compounds of thisdisclosure is that they are orally bioavailable and can be dosed orally.

Method for Treating Viral Infection

The present application provides a method for treating a Flaviviridaeviral infection comprising administering a therapeutically effectiveamount of a compound described herein or a pharmaceutically acceptablesalt, isotope, stereoisomer, mixture of stereoisomers, tautomer, esteror prodrug thereof, to a human subject in need thereof.

Also provided is a method for treating a Coronaviridae viral infectioncomprising administering a therapeutically effective amount of acompound described herein or a pharmaceutically acceptable salt,isotope, stereoisomer, mixture of stereoisomers, tautomer, ester orprodrug thereof, to a human subject in need thereof.

In one embodiment, the method of inhibiting or treating a diseasecomprises administering to an animal a composition comprising aneffective amount of one or more compounds of the invention and apharmaceutically acceptable carrier. The composition to be administeredmay further contain a secondary therapeutic agent as described above.

A method of the present application is particularly suitable for usewith humans, but may be used with other animals, particularly mammals,such as, for example, non-human primates, companion animals, farmanimals, laboratory animals, and wild and zoo animals.

A method of the present application is particularly useful to treatdiseases caused directly or indirectly by Flaviviridae virus since thecompounds of the present invention have inhibitory activity againstthose viruses. In some embodiments, therefore, a method of the presentinvention is used in inhibiting or treating diseases caused by aHepatitis C virus. In some embodiments, therefore, a method of thepresent invention is used in inhibiting or treating diseases caused by aHepatitis B virus. In an aspect, such a method is applied to a patientwith a disease caused by the viral infection such as dengue fever,yellow fever, hepatitis C, Japanese encephalitis, Kyasanur forestdisease, Murray valley encephalitis, St. Louis encephalitis, tick-borneencephalitis or West Nile encephalitis.

In some embodiments, a sustained virologic response is achieved at about12 weeks, at about 10 weeks, at about 8 weeks, at about 6 weeks, or atabout 4 weeks, or at about 4 months, or at about 5 months, or at about 6months, or at about 1 year, or at about 2 years.

A method of the present application is also particularly useful to treatdiseases caused directly or indirectly by Coronaviridae virus since thecompounds of the present invention have inhibitory activity againstthose viruses. In some embodiments, therefore, a method of the presentinvention is used in inhibiting or treating diseases caused by a SARScoronarirus. In an aspect, such a method is applied to a patient with adisease caused by the viral infection such as severe acute respiratorysyndrome (SARS), cancer, inflammation, obesity, acquired immunedeficiency syndrome (AIDS), or cirrhosis.

In another aspect, the compounds disclosed herein can be used fortreating cancer. In yet another aspect, the compounds disclosed hereincan be used for immunomodulation. In some embodiments, therefore, amethod of the present invention comprises adjusting an immune responseto a desired level, as in immunopotentiation, immunosuppression, orinduction of immunologic tolerance.

In some embodiments, the compound is administered for about 12 weeks. Infurther embodiments, the compound is administered for about 12 weeks orless, for about 10 weeks or less, for about 8 weeks or less, for about 6weeks or less, or for about 4 weeks or less. The compound may beadministered once daily, twice daily, once every other day, two times aweek, three times a week, four times a week, or five times a week.

EXAMPLES

The following examples are merely illustrative, and do not limit thisdisclosure in any way.

List of Abbreviations and Acronyms Abbreviation Meaning ° C. DegreeCelsius di-tBuXPhos2-Di-tert-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6′-triisopropyl-1,1′-biphenyl 2,6-lut. 2,6-lutidine MNBA2-Methyl-6-nitrobenzoic Anhydride 4AMS 4 Angstrom molecular sieves AcAcetyl app Apparent aq Aqueous atm atmosphere BINAP(2,2′-bis(diphenylphosphino)-1,1′-binaphthyl) Bn Benzyl Boctert-Butoxycarbonyl BOP Benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate br Broad Bu Butyl cat Catalytic cmCentimeter cod cyclooctadiene COMU(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)-dimethylamino-morpholino-carbenium hexafluorophosphate CP/Cp CyclopentylCPME Cyclopentyl methyl ether CSA Camphorsulfonic acid Cy/cHexCyclohexyl d Doublet DAST Diethylaminosulfur trifluoride dbadibenzylideneacetone DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DCCN,N′-dicyclohexylcarbodiimide DCE Dichloroethane DCM Dichloromethane ddDoublet of doublets DIPEA N,N-Diisopropylethylamine DMAP4-Dimethylaminopyridine DMF Dimethylformamide DMSO Dimethylsulfoxide dqDouble quartet EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide equivEquivalents ESI Electron Spray Ionization Et Ethyl g Grams HATU(Dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methaniminium hexafluorophosphate HDMSHexamethyldisilazane HOBT Hydroxybenzotriazole HPLC High-performanceliquid chromatography hrs/h Hours Hz Hertz IC₅₀ The half maximalinhibitory concentration i-Pr/iPr Isopropyl J Coupling constant KgKilogram LCMS Liquid chromatography-mass spectrometry LDA Lithiumdiisopropylamide M Molar m Multiplet m/z mass-to-charge ratio M+ Masspeak m-CPBA meta-Chloroperoxybenzoic acid Me Methyl mg Milligram MHzMegahertz min Minute mL Milliliter mM Millimolar mm Millimeter mmolMillimole mol Mole Ms Methanesulfonyl MW Microwave nM Nanomolar NMRNuclear magnetic resonance o-Tol o-Tolyl Ph Phenyl Pin Pinacolato PivPivaloyl ppm parts per million pTSA p-Toluenesulfonic acid py Pyridine qQuartet quant Quantitative rac Racemic Rf Retention factor RT/rt/r.t.Room temperature s Singlet or seconds sat. Saturated SEMCl2-Trimethylsilylethyoxymethyl chloride t Triplet TBAFTetra-n-butylammonium fluoride TBDMS/TBS tert-Butyldimethylsilyl tdTriplet of doublets TEA Triethylamine Tf Trifluoromethanesulfonyl TFATrifluoroacetic acid THF Tetrahydrofuran TLC Thin layer chromatographyTMS Trimethylsilyl Tr/tr Retention time Ts Tosyl UV Ultraviolet δChemical shift μL Microliter μM Micromolar μmol Micromole

For Examples 1 to 58, and Examples 93 to 107, unless otherwise stated,preparative HPLC was performed on a Gilson HPLC system, using an AgilentEclipse XDB/C18 7 micron, 250×21.2 mm semi-preparative column and anacetonitrile/water mobile phase at a flow rate of 20 mL/min.

For Examples 59 to 92, and Examples 108 to 112, unless otherwise stated,preparative HPLC was performed on a Shimadzu HPLC system, using a21.2×250 mm 10 micron C18 Phenomenex Gemini semi-preparative column andacetonitrile/water mobile phase at a flow rate of 20 mL/min.

Example 1(E)-(2R,3R)-7-(3-{(R)-1-[((S)-Hexahydro-pyridazine-3-carbonyl)-amino]-ethyl}-phenyl)-3-methoxy-2-methyl-hept-6-enoicacid (S)-2-methyl-1-((S)-1-methyl-2-oxo-ethylcarbamoyl)-propylester—Compound 1

Synthesis of Compound 1a:

A solution of1-((1R,5S)-10,10-dimethyl-3,3-dioxo-3lambda*6*-thia-4-aza-tricyclo[5.2.1.0*1,5*]dec-4-yl)-propan-1-one(3.95 g, 14.55 mmol) in toluene (50 mL) was prepared, then evaporated todryness. This process was repeated and the resulting white solid wasdissolved in anhydrous dichloromethane (16 mL). A small quantity ofcalcium hydride was added before adding tert-butyldimethylsilyltrifluoromethanesulfonate (3.83 mL, 14.5 mmol) and anhydroustriethylamine (2.33 mL, 16.7 mmol). The reaction mixture was stirred atroom temperature (RT) under a nitrogen atmosphere for 15 hours (“h”).The resulting solution was evaporated to yield a thick paste, which wasre-dissolved in anhydrous dichloromethane (15 mL) and added drop-wise toa stirred solution of 4-pentenal (2.69 g, 32.0 mmol) and titaniumtetrachloride (1 M in dichloromethane, 32 mL, 32 mmol) in anhydrousdichloromethane (20 mL) at −78° C., under a nitrogen atmosphere. Thereaction was stirred at −78° C. for 30 min before diluting withsaturated aqueous ammonium chloride solution (100 mL). The layers wereseparated and the aqueous layer was extracted with furtherdichloromethane (2×50 mL). The combined extract was dried over sodiumsulfate, filtered and evaporated to give a brown gum. This was purifiedby silica gel chromatography using iso-hexane/ethyl acetate 4:1 to yieldthe title compound (3.09 g, 60%) as a colorless gum.

Synthesis of Compound 1b:

A solution of 1a (250 mg, 0.703 mmol) in anhydrous dichloromethane (7mL) was prepared and trimethyloxonium tetrafluoroborate (208 mg, 1.406mmol) was added. The reaction mixture was stirred at room temperaturefor 15 h. The reaction mixture was treated with methanol (1 mL), then 2M hydrochloric acid (20 mL) and saturated brine (20 mL). The mixture wasextracted with ethyl acetate (3×15 mL) and the extract was dried oversodium sulfate, filtered and evaporated to give a yellow gum. The gumwas purified by silica gel chromatography using iso-hexanelethyl acetate4:1 to give the title compound (223 mg, 86%) as a colorless gum.

Synthesis of Compound 1c:

A solution of 2 M lithium hydroxide in water (5 mL) was added to astirred solution of 1b (223 mg, 0.60 mmol) in tetrahydrofuran (15 mL).The stirred mixture was heated to 60° C. for 15 h. The reaction mixturewas partially evaporated before adding 2 M hydrochloric acid (20 mL).The solution was extracted with ethyl acetate (3×15 mL). The extract wasdried over sodium sulfate, filtered and evaporated to give a yellow gum(209 mg). The gum was purified by silica gel chromatography usingiso-hexane/ethyl acetate 3:1 to yield the title compound (68 mg, 66%) asa yellow gum.

Synthesis of Compound 1d:

A solution of (S)-2-tert-butoxycarbonylamino-propionic acid (3.28 g,17.32 mmol) in acetonitrile (160 mL) was cooled to 0° C. before additionof N,N-diisopropylethylamine (12 mL, 69.3 mmol) then2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(6.57 g, 17.32 mmol). The reaction mixture was stirred at 0° C. for 20min and a solution of (S)-hexahydropyridazine-3-carboxylic acid2,2,2-trichloroethyl ester trifluoroacetic acid salt (preparationdescribed in Angew. Chem. Int. Ed. Engl. 1999, 38, 2443, 6.49 g, 17.3mmol) in acetonitrile (80 mL) was added. The reaction was allowed towarm to room temperature and was stirred for 15 h. The reaction mixturewas evaporated then re-dissolved in ethyl acetate (150 mL). The solutionwas washed with brine (150 mL). The brine was back extracted with ethylacetate (50 mL). The organic layers were combined, dried over sodiumsulfate, filtered and evaporated to give a dark oil. The oil waspurified by silica gel chromatography using iso-hexane/ethyl acetate 1:1to yield the title compound (6.88 g, 92%) as a colorless gum.

Synthesis of Compound 1e:

To 1d (1.5 g, 3.5 mmol) in anhydrous dichloromethane (30 mL) at 0° C.and under an atmosphere of nitrogen was added trimethylsilyltrifluoromethanesulfonate (941 μL, 5.2 mmol). The reaction mixture wasstirred at 0° C. for 1 h before adding N,N-diisopropylethylamine (2.2mL, 13.9 mmol) and then concentrated in vacuo to afford(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloroethyl ester. The resulting residue was dissolved inanhydrous acetonitrile (20 mL) and cooled to 0° C. before adding(S)-2-hydroxy-3-methyl-butyric acid (413 mg, 3.5 mmol),hydroxybenzotriazole monohydrate (796 mg, 5.2 mmol),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.02 g, 5.2 mmol) andN,N-diisopropylethylamine (1.7 mL, 10.4 mmol). The reaction mixture wasstirred at 0° C. for 90 min, warmed to room temperature and stirred for72 h and concentrated in vacuo. Ethyl acetate was added to the residue,which was then washed with saturated ammonium chloride and brine. Theorganic layer was dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by silica gelchromatography using iso-hexane/acetone 2:1 to give the title compound(1.1 g, 73% over 2 steps) as a white solid.

Synthesis of Compound 1f:

A stirred solution of 1e (830 mg, 1.92 mmol) in dichloromethane (20 mL)was prepared and 1c (34 mg, 2.00 mmol), N,N′-dicyclohexylcarbodiimide(594 mg, 2.88 mmol) and N,N-dimethylaminopyridine (23 mg, 0.192 mmol)were added. The reaction was stirred at room temperature for 18 h. Afurther quantity of 1c (34 mg, 2.00 mmol), N,N′-dicyclohexylcarbodiimide(594 mg, 2.88 mmol) and N,N-dimethylaminopyridine (23 mg, 0.192 mmol)were added and the reaction was stirred at room temperature for afurther 22 h. The reaction mixture was washed with water and the aqueouslayer was back extracted with dichloromethane (2×15 mL) and ethylacetate (2×15 mL). The organic layers were combined, dried over sodiumsulfate, filtered and evaporated to give a brown gum. The gum waspurified by silica gel chromatography using iso-hexanelethyl acetate 1:1then ethyl acetate to give the title compound (656 mg, 58%) as a whitesolid.

Synthesis of Compound 1g:

A solution of 1f (656 mg, 1.12 mmol) in tetrahydrofuran (30 mL) wasprepared and zinc powder (732 mg, 11.2 mmol) and aqueous ammoniumacetate solution (1 M, 8 mL, 8 mmol) were added. The reaction wasstirred at room temperature for 18 h. The mixture was filtered throughhyflo-supercel then acidified with ammonium chloride then further with 2M hydrochloric acid to pH 1. The mixture was extracted with ethylacetate (3×25 mL). The extract was dried over sodium sulfate, filteredand evaporated to give a solid which was azeotroped with toluene (40 mL)to give the title product (343 mg, 67%) as a colorless solid.

Synthesis of Compound 1h:

A solution of (R)-bromo-α-methylbenzylamine (1.023 g, 5.112 mmol) indichloromethane (20 mL) was subsequently treated with triethylamine (720μL, 5.112 mmol) and di-tert-butyl dicarbonate (1.784 g, 8.179 mmol).After overnight stirring at room temperature, the volatiles were removedin vacuo and the residue was purified by silica gel chromatography usinga 50 g Isolute cartridge eluted with a continuous gradient ofiso-hexane/diethyl ether 1:0 to 4:1 to afford the title compound (1.552g, 100%) as a white solid.

Synthesis of Compound 1i:

A solution of 1h (10.26 g, 0.0342 mol.) and tributyl(vinyl)stannane(32.5 g, 30 mL, 0.103 mol.) in toluene (175 mL) was purged with nitrogenfor 30 minutes before addition of bis(triphenylphosphine palladium(II)dichloride (2.38 g, 0.0034 mol). The stirred mixture was heated to 60°C. for 16 h before cooling to room temperature. The reaction mixture wasfiltered through hyflo-supercel then evaporated to give a dark coloredoil. The oil was purified by silica gel chromatography usingiso-hexane/ethyl acetate 19:1 to yield the title compound (6.95 g, 82%)as a yellow oil.

Synthesis of Compound 1j:

A solution of ii (6.95 g, 28.1 mmol.) in 1,4-dioxane (30 mL) wasprepared and a solution of hydrogen chloride in 1,4 dioxane (4M, 60 mL)was added. The reaction mixture was stirred at room temperature for 2 hthen evaporated to dryness. The resultant solid was re-dissolved intoluene and evaporated. The solid was triturated with diethyl ether,which was removed by decanting. The solid was then dried under vacuum togive the title compound (4.96 g, 96%) as an off-white solid.

Synthesis of Compound 1k:

A solution of 1g (343 mg, 0.752 mmol), 1j (138 mg, 0.752 mmol) andN,N-diisopropylethylamine (0.52 mL, 3.00 mmol) in acetonitrile (25 mL)was prepared and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate methanaminium (0.40 g, 1.05 mmol) was added. Themixture was left to stir at room temperature for 18 h. The reactionmixture was evaporated then re-dissolved in ethyl acetate (25 mL) andwater (25 mL). The layers were separated and the aqueous layer wasextracted with further ethyl acetate (2×20 mL). The combined organiclayers were dried over sodium sulfate, filtered and evaporated to give abrown gum. The gum was purified by silica gel chromatography usingiso-hexanelethyl acetate 1:3 to give the title compound (217 mg, 49%) asa colorless solid.

Synthesis of the Title Compound:

A stirred solution of 1k (217 mg, 0.37 mmol) in 1,2-dichloroethane (125mL) was prepared and Hoveyda-Grubbs 2^(nd) generation catalyst (23 mg,0.037 mmol) was added. The stirred solution was heated to reflux for 3h. Further Hoveyda-Grubbs 2^(nd) generation catalyst (46 mg, 0.074 mmol)was added and the reaction was heated to 85° C. for a further 2 h. Thecooled reaction mixture was treated with silica gel. The mixture wasevaporated then purified by silica gel chromatography using ethylacetate to give an off-white solid (177 mg). This was re-purified bysilica gel chromatography using a 5 g cartridge and ethyl acetate togive the title compound (67 mg, 32%) as a white solid. ¹H NMR (300 MHz,d₆-acetone) 0.93 (d, J=7.1 Hz, 3H), 0.95 (d, J=7.3 Hz, 3H), 1.15 (d,J=6.9 Hz, 3H), 1.34 (d, J=6.7 Hz, 3H), 1.43 (d, J=7.1 Hz, 3H), 1.47-1.69(m, 3H), 1.70-1.91 (m, 3H), 1.92-2.17 (m, 3H), 2.32-2.48 (m, 1H),3.02-3.13 (m, 1H), 3.35 (s, 3H), 3.40-3.65 (m, 3H), 4.27-4.43 (m, 2H),4.80 (d, J=5.3 Hz, 1H), 5.00-5.15 (m, 1H), 5.32-5.44 (m, 1H), 6.20-6.33(m, 1H), 6.42 (d, J=15.8 Hz, 1H), 7.10-7.32 (m, 3H), 7.34 (s, 1H), 7.87(d, J=8.0 Hz, 1H). LCMS (m/z) 557.3 [M+H], Tr=2.59 min.

Example 2 Compound 2

Synthesis of Compound 2a:

A solution of1-((1R,5S)-10,10-dimethyl-3,3-dioxo-3lambda*6*-thia-4-aza-tricyclo[5.2.1.0*1,5*]dec-4-yl)-propan-1-one(6.0 g, 22.1 mmol) in anhydrous dichloromethane (24 mL) was prepared andtert-butyldimethylsilyl trifluoromethanesulfonate (5.0 mL, 22.1 mmol)was added, followed by anhydrous triethylamine (3.54 mL, 25.4 mmol). Thereaction mixture was stirred at room temperature under a nitrogenatmosphere for 15 h. This gave a dark solution that was evaporated togive an oil. The oil was dissolved in anhydrous dichloromethane (22 mL)and the solution was added dropwise to a solution of crotonaldehyde(3.66 mL, 44.2 mmol) and titanium tetrachloride (1 M in dichloromethane,44.2 mL, 44.2 mmol) in dichloromethane (22 mL) at −78° C. under anitrogen atmosphere. The reaction mixture was stirred at −78° C. for 1h, before addition of ammonium chloride solution (30 mL). The stirredmixture was allowed to warm to room temperature before separating thelayers. The aqueous layer was extracted with dichloromethane (2×25 mL).The organic layers were combined, dried over sodium sulfate, filteredand evaporated to give a brown oil. The oil was purified by silica gelchromatography using iso-hexane/ethyl acetate 4:1 to yield the titlecompound (6.7 g, 89%) as a colorless solid.

Synthesis of Compound 2b:

A solution of 2a (4.15 g, 12.1 mmol) in anhydrous dichloromethane (80mL) was prepared and 1,8-bis(dimethylamino)naphthalene (7.78 g, 36.3mmol) was added followed by trimethyloxonium tetrafluoroborate (3.6 g,24.2 mmol). The reaction mixture was stirred at room temperature for 3h. The reaction mixture was treated with methanol (3 mL) and stirred for5 min before adding hydrochloric acid (2 M, 200 mL) and ethyl acetate(250 mL). The mixture was filtered to remove an insoluble solid and thelayers were separated. The aqueous layer was extracted with ethylacetate (2×100 mL). The organic layers were combined and washed withbrine, dried over sodium sulfate, filtered and evaporated to give thetitle compound (4.80 g, 100%) as a pale brown solid.

Synthesis of Compound 2c:

A solution of lithium hydroxide in water (2 M, 50 mL, 100 mmol) wasadded to a stirred solution of 2b (4.80 g, 12.1 mmol) in tetrahydrofuran(130 mL). The reaction mixture was heated to 60° C. for 15 h. Thereaction mixture was cooled to room temperature, before partiallyevaporating and adding hydrochloric acid (2 M, 150 mL). The mixture wasextracted with ethyl acetate (3×50 mL). The extract was dried oversodium sulfate, filtered and evaporated to give a brown oil (3.5 g). Theoil was purified by silica gel chromatography using iso-hexane/diethylether 1:1 to give the title compound (1.132 g, 59%) as a colorless oil.

Synthesis of Compound 2d:

Compound 2d was prepared in the same manner as compound 1f replacing icwith 2c (148 mg, 0.94 mmol) to afford the title compound (340 mg, 76%)as a white solid.

Synthesis of Compound 2e:

A solution of 6-bromo-3-chloro-isoquinoline (Frontier Scientific, Logan,Utah USA) (8.0 g, 33 mmol) and tributyl-(1-ethoxy-vinyl)-stannane (14.88g, 14 mL, 41.2 mmol) in toluene (100 mL) was degassed with nitrogen for30 min. Bis(triphenylphosphine)palladium(II) dichloride (1.16 g, 1.65mmol, 5 mol %) was added and the reaction mixture was heated at 60° C.for 20 h. The reaction mixture was cooled to room temperature, themixture was filtered and the filtrate was evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 20:1 to 10:1 to afford the title compound (7.1g, 92%) as a pale yellow solid.

Synthesis of Compound 2f:

A solution of 2e (7.1 g, 30 mmol) in 1,4-dioxane (60 mL) and 2 Mhydrochloric acid (30 mL) was stirred at room temperature for 30 min.The majority of the solvent was evaporated and the residue waspartitioned between ethyl acetate and water. The organic extracts werecombined, washed with water and brine, dried over sodium sulfate,filtered and evaporated. The residue was triturated with 5% ether iniso-hexane and the resulting solid was collected and dried to afford thetitle compound (6.0 g yield) as a white solid.

Synthesis of Compound 2g:

A solution of 2f (1.72 g, 8.3 mmol) in tetrahydrofuran (40 mL) wasstirred under nitrogen. Titanium (IV) ethoxide (3.8 g, 3.45 mL, 16.6mmol, tech. grade) was added followed by(R)-(+)-2-methyl-propanesulfinimide (1.11 g, 9.2 mmol) and the reactionmixture was stirred at 60° C. under nitrogen for 18 h. Additional(R)-(+)-2-methyl-propanesulfinimide (190 mg, 0.2 equiv) was added andthe reaction mixture was stirred at 65° C. for a further 2 h. Thereaction mixture was cooled to room temperature and ethyl acetate andbrine were added. The suspension was filtered through celite and thefilter pad was washed with ethyl acetate. The ethyl acetate layer wasseparated, washed with brine, dried over sodium sulfate, filtered andevaporated. The residue was purified by silica gel chromatography usinga gradient of iso-hexanes/ethyl acetate 7:3 to 3:7 to afford the titlecompound (2.2 g, 86%).

Synthesis of Compound 2h:

A mixture of (1S,2R)-(−)-cis-1-amino-2-Indanol (60 mg, 0.4 mmol),[Ru(p-cymen)Cl₂]₂ (122 mg, 0.2 mmol) and powdered 4 Å molecular sieves(2g) was suspended in anhydrous 2-propanol (9 mL) and stirred undernitrogen. The suspension was heated at 90° C. for 20 min. The reactionmixture was cooled to 40° C. and a solution of 2g (1.23 g, 4 mmol) in2-propanol (28 mL) was added followed by a solution of potassiumtert-butoxide (122 mg, 1.1 mmol) in 2-propanol (10 mL). The reactionmixture was stirred for 2 h at 40° C. and then allowed to cool. Themixture was poured directly onto a silica gel cartridge and eluted withethyl acetate to give, after evaporation, the title compound as a browngum (1.19 g, 96%).

Synthesis of Compound 2i:

A mixture of 2h (1.66 g, 2.11 mmol), tributyl(vinyl)tin (1.85 mL, 6.35mmol) and palladium tetrakis(triphenylphosphine) (488 mg, 0.42 mmol in1,4-dioxane (10.5 mL) was capped in a microwave vial. The reactionmixture was heated and stirred at 160° C. for 40 min in a microwavereactor. A second reaction was carried under identical scale andconditions and the reactions combined and evaporated. The residue waspurified by silica gel chromatography using a gradient of 25 to 100%ethyl acetate in iso-hexanes to afford the title compound as a brown gum(1g).

Synthesis of Compound 2j:

2i was suspended in 4 M HCl in 1,4-dioxane (17 mL, 68 mmol) and methanolwas added (34 mL). The reaction mixture was stirred for 90 min and thenevaporated to give the title compound as the di-HCl salt. The residuewas passed through an SCX cartridge eluting with methanol and thenmethanolic ammonia. The basic fraction was collected and evaporated togive the title compound as a beige solid (530 mg, 63% over 2 steps).

Synthesis of Compound 2k:

Compound 2k was prepared in the same manner as compound 1k, by firstreplacing 1g with(S)-1-{(S)-2-[(S)-2-((E)-(2R,3R)-3-methoxy-2-methyl-hex-4-enoyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (340 mg, 0.59 mmol) and following thesynthesis for compound 1g and then replacing(R)-1-(3-vinyl-phenyl)-ethylamine hydrochloride with 2j dihydrochloride(160 mg, 0.59 mmol) in the synthesis of compound 1k to give the titlecompound (240 mg, 66%, 2 steps) as a viscous light brown oil.

Synthesis of the Title Compound:

A solution of 2k (300 mg, 0.48 mmol) in toluene (161 mL) was degassedwith nitrogen for 10 min before adding Hoveyda Grubbs 2^(nd) generationcatalyst (61 mg, 0.1 mmol). The stirred solution was heated to 125° C.for 2 h before a further amount of Hoveyda Grubbs 2^(nd) generationcatalyst (61 mg, 0.1 mmol) was added and heating continued for 4 h. Thereaction was cooled to ambient temperature and concentrated to dryness.The residue was purified by silica gel chromatography using ethylacetate/acetone 1/0 then 3/1 followed by preparative TLC using ethylacetate/acetone 8/1 to yield the title compound (4 mg, 1%) as anoff-white solid. ¹H NMR (300 MHz, CDCl₃) 0.80-1.11 (m, 6H), 1.20-1.38(m, 3H), 1.42-1.70 (m, 6H), 1.67-1.88 (m, 2H), 1.89-2.25 (m, 3H),2.60-2.75 (m, 2H), 2.98-3.18 (m, 1H), 3.48 (s, 3H), 3.58-3.77 (m, 1H),4.36-4.67 (m, 2H), 4.91 (s, 1H), 5.18-5.43 (m, 1H), 5.61-5.76 (m, 1H),6.40-6.65 (m, 1H), 6.82-7.10 (m, 3H), 7.36-7.50 (m, 1H), 7.64 (s, 1H),7.85-8.01 (m, 1H), 8.18 (s, 1H), 9.16 (s, 1H). LCMS (m/z)=580.3 [M+H],Tr=1.50 min.

Example 3 Compound 3

Synthesis of Compound 3a:

Compound 3a was prepared in the same manner as compound 1f by replacingic with 3-butenoic acid to afford the title compound (397 mg, 66%) as awhite solid.

Synthesis of Compound 3b:

To a solution of 3a (390 mg, 0.78 mmol) in tetrahydrofuran (17 mL) andwater (9 mL) was added powdered zinc (1.11 g, 17.1 mmol) followed byammonium acetate (899 mg, 11.7 mmol). The reaction mixture was stirredat room temperature for 15 h. The reaction was filtered through Celite,the filtrate was acidified to pH 2 with concentrated hydrochloric acidand extracted twice with ethyl acetate. The organic layers were driedthrough a hydrophobic frit and concentrated in vacuo, followed byco-evaporation from toluene (2×). The ensuing residue was dissolved inanhydrous acetonitrile (15 mL) and 2j (211 mg, 0.78 mmol) was addedfollowed by hydroxybenzotriazole monohydrate (119 mg, 0.78 mmol),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (210 mg, 1.1 mmol). Thereaction mixture was stirred at room temperature for 16 h. Silica gelwas added to the reaction mixture and concentrated in vacuo. The residuewas purified by silica gel chromatography using iso-hexane/ethyl acetate1/1 then ethyl acetate to give the title compound (276 mg, 57%, 2 steps)as a white solid.

Synthesis of the Title Compound:

To a refluxing solution of Hoveyda Grubbs 2^(nd) generation catalyst (85mg, 0.14 mmol) and 2,6-dichlorobenzoquinone (8 mg, 0.05 mmol) inanhydrous toluene (150 mL) was added a solution of 3b (250 mg, 0.45mmol) in toluene/dichloroethane (1:1, 9 mL) over 1 h. Following 1 h atreflux, a further amount of Hoveyda Grubbs 2^(nd) generation catalyst(42 mg, 0.07 mmol) was added and heating continued for 3 h. The reactionwas cooled to room temperature and evaporated to dryness. The residuewas purified by silica gel chromatography using ethyl acetate then ethylacetate/acetone 3/1 to yield a brown foam. This was purified further bypreparative TLC using ethyl acetate/acetone 8/1 (2×) to yield the titlecompound (2 mg, 1%) as a white solid. ¹H NMR (300 MHz, CDCl₃) 0.73-1.20(m, 9H), 1.40-2.08 (m, 5H), 2.15-2.61 (m, 4H), 3.14-4.15 (m, 6H),4.78-5.63 (m, 3H), 6.67-7.03 (m, 2H), 7.38-7.58 (m, 2H), 7.83-8.04 (m,2H), 9.17 (s, 1H). LCMS (m/z)=522.2 [M+H], Tr=1.33 min.

Example 4 Compound 4

Synthesis of Compound 4a:

To a solution of (S)-4-benzyl-2-oxazolidinone (3.85 g, 21.7 mmol) intetrahydrofuran (10 mL) at −78° C., was added 2.5 M butyllithium inhexanes (10 mL, 25 mmol) and the mixture stirred for 5 min. Valericanhydride (5.15 mL, 26.1 mmol) was then added and the solution stirredat −78° C. for 50 min before addition of brine (10 mL). The mixture wasthen allowed to warm to room temperature when more brine (20 mL) wasadded. The organics were extracted with ethyl acetate (2×50 mL) and thecombined extract was dried over sodium sulfate, filtered and evaporatedto give a crude oil. This was purified by silica gel chromatographyusing iso-hexane/ethyl acetate 9:1 to yield the title compound (5.24 g,92%).

Synthesis of Compound 4b:

A solution of 4a (5.24 g, 20.0 mmol) in tetrahydrofuran (50 mL) wascooled to −78° C. 1 M sodium hexamethyldisilazane in tetrahydrofuran(32.1 mL, 32 mmol) was added dropwise and the internal temperature keptbelow −70° C. Upon complete addition, stirring was continued at −78° C.for 1 h. After this time allyl bromide (6.98 mL, 80.2 mmol) was addeddropwise and the internal temperature kept between −55° C. to −40° C.Upon complete addition stirring was continued between −55° C. and −40°C. for 3 h, after which saturated ammonium chloride (20 mL) was addedand the mixture allowed to warm to room temperature. The organics wereextracted with ethyl acetate (2×100 mL), washed with brine, dried oversodium sulfate and evaporated to give a crude yellow oil. This waspurified by silica gel chromatography using iso-hexane/ethyl acetate 1:0then 95:5 then 9:1 to yield the title compound (2.95 g, 49%).

Synthesis of Compound 4c:

A solution of 4b (2.95 g, 9.79 mmol) in tetrahydrofuran (72 mL) andwater (36 mL) was cooled to 0° C. 30% aqueous hydrogen peroxide (4.72mL, 48.9 mmol) was added followed by a solution of lithium hydroxide(470 mg, 19.6 mmol) in water (7.2 mL). The mixture was stirred at 0° C.and after 3.42 h a solution of sodium sulphite (11.3 g, 90 mmol) inwater (36 mL) was added and the solution stirred at 0° C. for 15 min.The solution was treated with saturated sodium hydrogen carbonate untilpH=9-10, then the aqueous phase was washed with ethyl acetate (3×100 mL)then acidified to pH=1-2 with 1 M sulphuric acid and extracted withdiethyl ether (3×100 mL). The diethyl ether extracts were dried oversodium sulfate, filtered and evaporated to give the title compound (649mg, 47%) as a colorless oil.

Synthesis of Compound 4d:

To a solution of 1e (790 mg, 1.82 mmol) in dichloromethane (9 mL) atroom temperature, was added 4c (388 mg, 2.19 mmol),N,N′-dicyclohexylcarbodiimide (564 mg, 2.74 mmol) and4-dimethylaminopyridine (223 mg, 1.82 mmol) and the reaction mixturestirred at room temperature for 3 h. After this time the reactionmixture was diluted with dichloromethane (50 mL) and washed with asaturated ammonium chloride solution. The aqueous phase was re-extractedwith dichloromethane (2×50 mL) and the combined organics washed withbrine (100 mL), dried over magnesium sulfate and evaporated to give awhite residue. This was purified by silica gel chromatography usingiso-hexane/ethyl acetate 1:0 to 1:1 to yield the title compound (916 mg,90%).

Synthesis of Compound 4e:

A solution of 4d (916 mg, 1.65 mmol) in tetrahydrofuran (41 mL) wasprepared and powdered zinc (2.37 g, 36.2 mmol) was added followed by asolution of ammonium acetate (1.90 g, 24.7 mmol) in water (10.2 mL). Thereaction mixture was stirred at room temperature for 24 h. The reactionwas filtered through hyflo-supercel washing through with ethyl acetateand saturated aqueous potassium hydrogen sulfate. The mixture wastreated with hydrochloric acid (1 M, 5 mL) and the layers wereseparated. The aqueous layer was extracted with ethyl acetate (2×100mL). The organic layers were combined, washed with brine, dried oversodium sulfate, filtered and evaporated to give a colorless gum. Theresidue was azeotroped with toluene (3×100 mL) to give the titlecompound (675 mg, 89%) as a white solid.

Synthesis of Compound 4f:

4e (368 mg, 0.87 mmol) was dissolved in acetonitrile (5 mL) and 2j (180mg, 0.77 mmol) was added followed by N,N-diisopropylethylamine (535 μL,3.07 mmol) and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (409 mg, 1.08 mmol). The reactionmixture was stirred at room temperature for 72 h. To the reactionmixture was added 2 M hydrochloric acid (15 mL) and the mixture wasconcentrated in vacuo. The residue was partitioned between water (100mL) and ethyl acetate (100 mL). The aqueous layer was separated andre-extracted with ethyl acetate (100 mL). The organic layers werecombined, dried over sodium sulfate, filtered and evaporated to give acrude residue. The residue was purified by silica gel chromatographyusing ethyl acetate to give the title compound (440 mg, 95%) as anoff-white solid.

Synthesis of the Title Compound:

A solution of 4f (440 mg, 0.73 mmol) in toluene (242 mL) was degassedwith nitrogen for 15 min then Hoveyda-Grubbs 2^(nd) generation catalyst(91 mg, 0.15 mmol) was added and the reaction mixture heated at 100-110°C. for 2.5 h. After this time more Hoveyda-Grubbs 2^(nd) generationcatalyst (91 mg, 0.15 mmol) was added and the reaction mixture heated at110° C. for 1 h and allowed to cool to room temperature. The residue waspurified by silica gel chromatography using a gradient ofiso-hexane/ethyl acetate 1:1 to neat ethyl acetate. Impure product (160mg) was further purified by silica gel chromatography using a gradientof dichloromethane/acetone 99:1 to 4:1. Pure product (4.4 mg) wasobtained along with impure fractions which were further purified byreverse phase preparative HPLC where pure product (22.6 mg) wascollected. After purification the title compound was collected (27 mgtotal, 6%) as a white solid. ¹H NMR (300 MHz, CD₃OD) 0.93-1.11 (m, 11H),1.24-1.47 (m, 6H), 1.63-1.82 (m, 3H), 1.84-2.01 (m, 3H), 2.41-2.72 (m,2H), 2.82 (td, J=13.1, 3.1 Hz, 1H), 3.59-3.70 (m, 1H), 4.46 (dd, J=9.8,3.3 Hz, 1H), 4.70-4.79 (m, 2H), 5.17 (q, J=6.9 Hz, 1H), 5.83-5.96 (m,1H), 6.70 (d, J=16.5 Hz, 1H), 6.85-6.99 (m, 1H), 7.59 (dd, J=8.5, 1.6Hz, 1H), 7.71 (s, 1H), 7.96 (s, 1H), 8.03 (d, J=8.5 Hz, 1H), 8.54 (br d,J=8.9 Hz, 1H), 9.10 (s, 1H). LCMS (m/z) 578.3 [M+H], Tr=1.67 min.

Example 5 Compound 5

Synthesis of Compound 5a:

To (S)-4-benzyl-3-((R)-2-methyl-pent-4-enoyl)-oxazolidin-2-one (Preparedas in JACS 1990, 112 (8) 2998-3017, 2.5 g, 9.16 mmol) in ethanol (13 mL)and water (1.5 mL) at room temperature was added rhodium trichloridehydrate (48 mg, 0.23 mmol). The reaction was heated to 80° C. for 8 h,cooled and concentrated in vacuo. To the ensuing residue was added waterand extracted with dichloromethane. The organic layer was dried througha hydrophobic frit and concentrated in vacuo to yield the title compound(2.2 g, 88%) as a brown oil.

Synthesis of Compound 5b:

To 5a (2.2 g, 8.1 mmol) in tetrahydrofuran (60 mL) and water (20 mL) at0° C. was added lithium hydroxide (16 mL, 2.0 M solution in water, 32.2mmol) and hydrogen peroxide (6.6 mL, 30% aqueous solution, 64.5 mmol).The reaction was stirred at 0° C. for 16 h and then quenched by carefuladdition of aqueous sodium thiosulfate. The reaction mixture wasconcentrated to remove the tetrahydrofuran and then washed twice withdichloromethane. The aqueous layer was acidified to pH 1 with 2 Mhydrochloric acid and extracted twice with diethyl ether. The combinedorganic layers were dried through a hydrophobic frit and concentrated invacuo. The residue was purified by silica gel chromatography usingiso-hexane/ethyl acetate 5/1 to afford the title compound (530 mg, 57%)as a clear oil.

Synthesis of Compound 5c:

Compound 5c was prepared in the same manner as 1f replacing 1c with 5b(250 mg, 2.2 mmol), to afford the title compound (750 mg, 77%) as awhite foam.

Synthesis of Compound 5d:

To a solution of 5c (750 mg, 1.42 mmol) in tetrahydrofuran (35 mL) andwater (20 mL) was added powdered zinc (2.03 g, 31.2 mmol) followed byammonium acetate (1.64 g, 21.3 mmol). The reaction mixture was stirredat room temperature for 15 h. The reaction was filtered through Celite,the filtrate was acidified to pH 2 with concentrated hydrochloric acidand extracted twice with ethyl acetate. The organic layers were driedthrough a hydrophobic frit and concentrated in vacuo, followed byco-evaporation from toluene (2×). The ensuing residue was dissolved inanhydrous acetonitrile (15 mL) and 2j (385 mg, 1.4 mmol) was addedfollowed by N,N-diisopropylethylamine (743 μL, 4.3 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (755 mg, 2.0 mmol). The reactionmixture was stirred at room temperature for 3 h. Silica gel was added tothe reaction mixture and concentrated in vacuo. The residue was purifiedby silica gel chromatography using ethyl acetate to give the titlecompound (580 mg, 71%, 2 steps) as a clear viscous oil.

Synthesis of the Title Compound:

A solution of 5d (250 mg, 0.43 mmol) in toluene (144 mL) was degassedwith nitrogen for 30 min before warming to 70° C. and adding HoveydaGrubbs 2^(nd) generation catalyst (54 mg, 0.09 mmol). The stirredsolution was heated to 125° C. for 45 min. A further amount of HoveydaGrubbs 2^(nd) generation catalyst (50 mg, 0.08 mmol) was added andheating continued for 2 h. Another portion of Hoveyda Grubbs 2^(nd)generation catalyst (27 mg, 0.04 mmol) was added and heating continuedfor 3 h. The reaction was cooled to room temperature and potassiumisocyanoacetate (120 mg) in methanol (2 mL) was added. After stirringfor 1 h, the reaction mixture was concentrated to approximately half itsoriginal volume, treated with silica gel and then evaporated to dryness.The residue was purified by silica gel chromatography using ethylacetate/acetone 1/0 then 5/1 to yield a white solid. This was purifiedfurther by reverse phase preparative HPLC to yield the title compound (2mg, 1%) as a white solid. ¹H NMR (300 MHz, d₄-MeOH, 2:1 diastereomericmix, major diastereomer peaks quoted) 0.99 (d, J=6.7 Hz, 3H), 1.01 (d,J=6.7 Hz, 3H), 1.47 (d, J=6.9 Hz, 3H), 1.56-1.64 (m, 6H), 1.65-1.79 (m,1H), 1.83-2.00 (m, 3H), 2.09-2.25 (m, 1H), 2.73-2.87 (m, 1H), 3.41-3.52(m, 1H), 3.63-3.71 (m, 1H), 4.38-4.48 (m, 1H), 5.12 (d, J=8.5 Hz, 1H),5.18 (q, J=6.7 Hz, 1H), 5.82 (q, J=7.1 Hz, 1H), 6.47 (dd, J=16.0, 6.3Hz, 1H), 6.67 (dd, J=16.0, 1.0 Hz, 1H), 7.60-7.67 (m, 2H), 7.75 (s, 1H),8.06 (d, J=8.7 Hz, 1H), 9.14 (s, 1H). LCMS (m/z)=536.2 [M+H], Tr=1.30min.

Example 6 Compound 6

Synthesis of Compound 6a:

A solution of (R)-bromo-α-methylbenzylamine (2.0 g, 10 mmol) indichloromethane (20 mL) was treated with a solution of di-tert-butyldicarbonate (2.4 g, 11 mmol) in dichloromethane (20 mL) and the reactionmixture was stirred at room temperature for 1 h. The solution was washedwith 2 M hydrochloric acid, water and brine, dried over sodium sulfate,filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanelethyl acetate 20:1 to 6:1to afford the title compound (2.51 g, 84%) as a white solid.

Synthesis of Compound 6b:

A mixture of 6a (900 mg, 3 mmol), 4-(2methoxycarbonylethyl)phenylboronicacid (624 mg, 3 mmol) in 1,2-dimethoxyethane (10 mL) and potassiumcarbonate (828 mg, 6 mmol) in water (2 mL), was stirred at roomtemperature. Tetrakis(triphenylphosphine)palladium(0) (172 mg, 0.15mmol) was added and the reaction mixture was heated at 100° C. in amicrowave reactor for 1 hour. The organic layer was separated. Theorganic layer was filtered through a hydrophobic frit and the filtratewas evaporated. The residue was purified by silica gel chromatographyusing a gradient of iso-hexanes/ethyl acetate 20:1 to 5:1 to afford thetitle compound (958 mg, 83%) as a white solid.

Synthesis of Compound 6c:

A solution of 6b (958 mg, 2.5 mmol) in tetrahydrofuran (15 mL) wasstirred at 5° C. A solution of lithium hydroxide hydrate (300 mg, 5mmol) in water (5 mL) was added and the reaction mixture was stirred at5° C. for 1 hour and then at room temperature for 1 hour. The majorityof the solvent was evaporated and water was added and 2 M hydrochloricacid was added to adjust the pH of the solution to pH 2. The mixture wasextracted with ethyl acetate. The organic extracts were combined, washedwith water and brine. The organic layer was filtered through ahydrophobic frit and the filtrate was evaporated to afford the titlecompound (883 mg, 96%) as a white solid.

Synthesis of Compound 6d:

A solution of 6c (1.6 g, 4.3 mmol), 1e (1.86 g, 4.3 mmol) indichloromethane (60 mL) was stirred at room temperature under nitrogen.N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (1.25 g,6.5 mmol) and 4-dimethylaminopyridine (525 mg, 4.3 mmol) was added andthe reaction mixture was stirred at room temperature under nitrogen for24 h. The reaction mixture was diluted with dichloromethane and thesolution was washed with an aqueous citric acid solution (pH 2-3), waterand brine. The organic layer was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using iso-hexanes/ethyl acetate 6:4 followed by silicagel chromatography using iso-hexanes/ethyl acetate 6:4 to afford thetitle compound (2.0 g, 60%) as a white foam.

Synthesis of Compound 6e:

A solution of 6d (1.96 g, 2.5 mmol) in tetrahydrofuran (50 mL) wasstirred at room temperature under nitrogen. Zinc dust (3.58 g, 55 mmol)was added followed by a solution of ammonium acetate (2.9 g, 37.5 mmol)in water (25 mL) and the reaction mixture was stirred at roomtemperature under nitrogen for 24 hours. The reaction mixture wasfiltered through Celite and the filter pad was washed with ethyl acetateand 2 M hydrochloric acid. The filtrate was acidified to pH 2 with 2 Mhydrochloric acid and sodium chloride was added to saturate the aqueouslayer. The mixture was extracted with ethyl acetate (3×). The organicextracts were combined, washed with water and brine, dried overanhydrous magnesium sulfate and the solvent was evaporated. The residuewas co-evaporated with ethyl acetate (3×), toluene (4×) and dried toafford the title compound (1.56 g, 95%) as a white solid.

Synthesis of the Title Compound:

A mixture of 6e (850 mg, 1.3 mmol) in 4M HCl in 1,4-dioxane (12 mL) wasstirred at room temperature under nitrogen for 1 h. The solvent wasevaporated and the residue was triturated with diethyl ether (3×). Theresulting solid was collected, washed with diethyl ether and dried toafford(S)-1-[(S)-2-((S)-2-{3-[3′-((R)-1-amino-ethyl)-biphenyl-4-yl]-propionyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride (1.3 mmol). A solution of(S)-1-[(S)-2-((S)-2-{3-[3′-((R)-1-amino-ethyl)-biphenyl-4-yl]-propionyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride (1.3 mmol) and N,N-diisopropylethylamine (671 mg, 0.9mL, 5.2 mmol) in dichloromethane (1300 mL) was stirred at 0° C. undernitrogen. 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (988 mg, 2.6 mmol) was added and thereaction mixture was stirred at room temperature for 20 h. The solventwas evaporated and the residue was purified by silica gel chromatographyusing a gradient of iso-hexanes/acetone 7:3 to 1:1 followed by silicagel chromatography using ethyl acetate. The resulting solid wastriturated with ether and the solid was collected and dried to affordthe title compound (395 mg, 57%) as a white solid. ¹H NMR (300 MHz,CD₃OD) δ0.94 (d, J=6.7 Hz, 3H), 1.00 (d, J=6.7 Hz, 3H), 1.31 (d, J=7.1Hz, 3H), 1.54 (d, J=6.9 Hz, 3H), 1.85-2.20 (m, 5H), 2.65-3.10 (m, 5H),3.53-3.59 (m, 1H), 4.38-4.25 (m, 1H), 4.64 (d, J=12.5 Hz, 1H), 4.72 (d,J=9.6 Hz, 1H), 5.05 (q, J=6.9 Hz, 1H), 5.24 (q, J=7.1 Hz, 1H), 4.82-4.90(m, 2H), 7.23-7.30 (m, 3H), 7.37-7.55 (m, 5H). LCMS (m/z) 535.2 [M+H],Tr=5.11 min.

Example 7(E)-(2R,5S,11S,14S,17R,18R)-18-Hydroxy-14-isopropyl-2,11,17-trimethyl-15-oxa-3,9,12,28-tetraaza-tricyclo[21.3.1.1*5,9*]octacosa-1(26),21,23(27),24-tetraene4,10,13,16-tetraone—Compound 7

Synthesis of Compound 7a:

A solution 1a (12.0 g, 0.034 mol) in anhydrous dichloromethane (520 mL)was cooled to 0° C. before addition of pyridine (5.5 mL, 0.068 mol),then trifluoromethanesulfonic acid tert-butyldimethylsilyl ester (9 mL,0.039 mol). The reaction was stirred at 0° C. for 15 min then allowed towarm to room temperature for 1.5 h. The reaction mixture was quenchedwith saturated aqueous sodium bicarbonate solution (400 mL). The aqueouslayer was back extracted with dichloromethane (200 mL). The organiclayers were combined and washed with brine (200 mL) and then 2 Mhydrochloric acid (200 mL). The solution was dried over sodium sulfate,filtered and evaporated to give the title product (15.29 g, 96%) as awhite solid.

Synthesis of Compound 7b:

A solution of 7a (15.29 g, 0.0325 mol) in tetrahydrofuran (300 mL) wasprepared and an aqueous solution of lithium hydroxide (2 M, 120 mL) wasadded. The stirred mixture was heated to 60° C. for 16 h. The reactionmixture was cooled to room temperature then treated with 2 Mhydrochloric acid (250 mL). The layers were separated and the aqueouslayer was extracted with ethyl acetate (2×200 mL). The organic layerswere combined, dried over sodium sulfate, filtered and evaporated togive an off-white solid. The solid was purified by silica gelchromatography using diethyl ether/iso-hexane 3:7 to yield the titleproduct (7.18 g, 81%) as a colorless gum.

Synthesis of Compound 7c:

To 1e (510 mg, 1.2 mmol) in anhydrous dichloromethane (4 mL) at roomtemperature and under an atmosphere of nitrogen was added 7b (385 mg,1.41 mmol), dicyclohexylcarbodiimide (364 mg, 1.8 mmol) and4-N,N-dimethylaminopyridine (142 mg, 1.2 mmol). The white suspension wasstirred at room temperature for 20 h before adding silica gel andconcentrating in vacuo. The residue was purified by silica gelchromatography using iso-hexanelethyl acetate 2/1 to afford the titlecompound (700 mg, 86%) as a viscous, clear oil.

Synthesis of Compound 7d:

To a solution of 7c (700 mg, 1.02 mmol) in tetrahydrofuran (22 mL) andwater (13 mL) was added powdered zinc (1.46 g, 22.4 mmol) followed byammonium acetate (1.2 g, 15.3 mmol). The reaction mixture was stirred atroom temperature for 15 h. The reaction was filtered through Celite andconcentrated in vacuo to remove the tetrahydrofuran. The ensuingsolution was acidified to pH 1-2 with concentrated hydrochloric acid andextracted with dichloromethane (2×). The organic layers were driedthrough a hydrophobic frit and concentrated in vacuo, followed byco-evaporation from toluene (2×). The ensuing colorless viscous oil wasdissolved in anhydrous acetonitrile (8 mL) and 1j (187 mg, 1.0 mmol) wasadded followed by N,N-diisopropylethylamine (900 μL, 5.1 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (542 mg, 1.4 mmol) The reactionmixture was stirred at room temperature for 15 h. Silica gel was addedto the reaction mixture and concentrated in vacuo. The residue waspurified by silica gel chromatography using iso-hexane/ethyl acetate 1/1to give the title compound (450 mg, 64%) as an off-white solid.

Synthesis of the Title Compound:

A solution of 7d (450 mg, 0.66 mmol) in 1,2-dichloroethane (219 mL) wasdegassed with nitrogen for 15 min before adding Hoveyda Grubbs 2^(nd)generation catalyst (41 mg, 0.07 mmol). The stirred solution was heatedto 80° C. for 2 h. The reaction was cooled, concentrated toapproximately 20 mL, treated with silica gel and then evaporated todryness. The residue was purified by silica gel chromatography usingiso-hexane/ethyl acetate 1/1 then 1/3 to yield(E)-(2R,5S,11S,14S,17R,18R)-18-(tert-butyl-dimethyl-silanyloxy)-14-isopropyl-2,11,17-trimethyl-15-oxa-3,9,12,28-tetraaza-tricyclo[21.3.1.1*5,9*]octacosa-1(26),21,23(27),24-tetraene-4,10,13,16-tetraone(138 mg) as a brown foam. The foam (80 mg, 0.12 mmol) was dissolved inanhydrous tetrahydrofuran (6.5 mL) and transferred to a polypropylenevial. At ambient temperature hydrogen fluoride*pyridine (102 L, 1.2mmol) was added to the reaction mixture followed by stirring for 1 h. Anadditional amount of hydrogen fluoride*pyridine (600 μL, 7.1 mmol) wasadded, followed by stirring for 1 h. More hydrogen fluoride*pyridine(700 μL, 8.2 mmol) was added and after 1 h the reaction was quenched bythe slow addition of the reaction mixture to a stirred solution of ethylacetate/saturated aqueous ammonium chloride. The organic layer wasseparated and the aqueous layer further extracted with ethyl acetate(2×). The combined organic layers were dried through a hydrophobic fritand concentrated in vacuo. The residue was purified by silica gelchromatography using ethyl acetate/acetone 8/1 to afford the titlecompound (33 mg, 16%, 2 steps) as a white solid. ¹H NMR (500 MHz, CD₃CN)0.92 (d, J=6.7 Hz, 3H), 0.97 (d, J=6.7 Hz, 3H), 1.26 (d, J=7.3 Hz, 3H),1.31 (d, J=6.7 Hz, 3H), 1.41 (d, J=6.7 Hz, 3H), 1.43-1.63 (m, 4H),1.78-1.89 (m, 2H), 2.03-2.09 (m, 1H), 2.40-2.49 (m, 2H), 2.50-2.56 (m,1H), 2.65-2.72 (m, 1H), 3.32 (td, J=11.0, 2.4 Hz, 1H), 3.49-3.61 (m,1H), 4.07 (d, J=12.2 Hz, 1H), 4.31 (d, J=12.8 Hz, 1H), 4.74 (d, J=5.5Hz, 1H), 4.97-5.04 (m, 1H), 5.18-5.25 (m, 1H), 6.27-6.34 (m, 1H), 6.38(d, J=15.9 Hz, 1H), 6.96-7.31 (m, 6H). LCMS (m/z)=543.2 [M+H], Tr=2.27min.

Example 8(E)-(2R,5S,11S,14S)-14-Isopropyl-2,11-dimethyl-15-oxa-3,9,12,28-tetraaza-tricyclo[21.3.1.1*5,9*]octacosa-1(27),21,23,25-tetraene-4,10,13,16-tetraone—Compound8

Synthesis of Compound 8a:

To a solution of 1e (791 mg, 1.83 mmol) in dichloromethane (9 mL) atroom temperature, was added hept-6-enoic acid (0.296 mL, 2.19 mmol),N,N′-dicyclohexylcarbodiimide (564 mg, 2.74 mmol) and4-N,N-dimethylaminopyridine (223 mg, 1.82 mmol) and the reaction mixturewas stirred at room temperature for 2 h. After this time the reactionmixture was diluted with dichloromethane (50 mL) and washed withsaturated aqueous ammonium chloride solution. The aqueous phase wasre-extracted with dichloromethane (2×50 mL) and the combined organicswashed with brine (100 mL) and dried over magnesium sulfate andevaporated to give a white residue. The residue was purified by silicagel chromatography using a gradient of iso-hexanelethyl acetate 1:1 toneat ethyl acetate to yield the title compound (774 mg, 81%).

Synthesis of Compound 8b:

A solution of 8a (650 mg, 1.20 mmol) in tetrahydrofuran (29 mL) wasprepared and powdered zinc (1.72 g, 26.3 mmol) was added followed by asolution of ammonium acetate (1.38 g, 18.0 mmol) in water (7.0 mL). Thereaction mixture was stirred at room temperature for 72 h. The reactionwas filtered through hyflo-supercel washing through with ethyl acetateand saturated aqueous potassium hydrogen sulfate. The mixture wastreated with hydrochloric acid (1 M, 3 mL) and the layers wereseparated. The aqueous layer was extracted with ethyl acetate (2×100mL). The organic layers were combined, washed with brine, filtered andevaporated to give a colorless gum. The residue was azeotroped withtoluene (3×100 mL) to give the title compound (492 mg, quantitativeyield) as a white solid.

Synthesis of Compound 8c:

To 8b (257 mg, 0.63 mmol) dissolved in acetonitrile (2 mL) was added 1j(122 mg, 0.83 mmol) followed by N,N-diisopropylethylamine (435 μL, 2.5mmol) and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (333 mg, 0.88 mmol). The reactionmixture was stirred at room temperature for 48 h. The mixture hadevaporated to dryness over the 48 hours so this residue was partitionedbetween water (100 mL) and ethyl acetate (100 mL). The aqueous layer wasseparated and re-extracted with ethyl acetate (100 mL). The organiclayers were combined, dried over magnesium sulfate, filtered andevaporated to give a crude residue. The residue was purified by silicagel chromatography using a gradient of iso-hexanelethyl acetate 7:3 to1:1 to neat ethyl acetate to give the title compound (194 mg, 57%).

Synthesis of the Title Compound:

A solution of 8c (194 mg, 0.36 mmol) in 1,2-dichloroethane was degassedwith nitrogen for 15 min then Hoveyda-Grubbs 2nd generation catalyst (23mg, 0.04 mmol) was added and the reaction mixture heated at 80° C. for2.3 h. After this time the reaction mixture was allowed to cool to roomtemperature and concentrated in vacuo. The residue was purified bysilica gel chromatography using a gradient of iso-hexane/ethyl acetate1:9 to neat ethyl acetate. Impure product (74 mg) was obtained which wasfurther purified by preparatory reverse phase HPLC where the titlecompound was collected (28 mg, 15%) as a white solid. ¹H NMR (300 MHz,CD₃OD). 0.96 (d, J=6.9 Hz, 3H), 0.99 (d, J=6.9 Hz, 3H), 1.41-1.51 (m,8H), 1.59-1.82 (m, 4H), 1.85-1.96 (m, 2H), 2.02-2.26 (m, 3H), 2.32-2.57(m, 2H), 2.80 (td, J=12.9, 2.2 Hz, 1H), 3.50 (dd, J=11.0, 2.5 Hz, 1H),4.39 (br d, J=13.2 Hz, 1H), 4.76 (d, J=8.0 Hz, 1H), 4.99 (q, J=6.9 Hz,1H), 5.46 (q, J=7.1 Hz, 1H), 6.19 (m, 1H), 6.40 (d, J=15.8 Hz, 1H),7.11-7.17 (m, 1H), 7.20-7.30 (m, 3H). LCMS (m/z) 513.2 [M+H], Tr=2.58min.

Example 9 Compound 9

To a solution of compound 4 (16.6 mg, 0.0287 mmol) in ethyl acetate (10mL) was added 10% palladium on carbon (5 mg) and stirred under ahydrogen atmosphere for 1.5 h. Then additional 10% palladium on carbon(5 mg) was added and stirring continued under a hydrogen atmosphere for24 h. The reaction mixture was filtered through Celite to remove thecatalyst and evaporated then purified by preparatory reverse phasepreparative HPLC to yield the title compound (5.3 mg, 32%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) 0.89-1.05 (m, 9H), 1.29-1.46 (m, 2H),1.59 (dd, J=7.1, 1.6 Hz, 6H), 1.65-1.99 (m, 7H), 2.09-2.18 (m, 1H),2.43-2.58 (m, 1H), 2.74-3.06 (m, 3H), 3.57-3.68 (m, 1H), 4.43 (dd,J=13.4, 2.7 Hz, 1H), 4.59 (br s, 3H), 4.66 (d, J=7.4 Hz, 1H), 5.15 (q,J=6.9 Hz, 1H), 5.66-5.78 (m, 1H), 7.58 (dd, J=8.5, 1.6 Hz, 1H), 7.69 (s,1H), 7.70 (s, 1H), 8.03 (d, J=8.5 Hz, 1H), 8.40 (br d, J=8.7 Hz, 1H),9.08 (s, 1H). LCMS (m/z) 580.3 [M+H], Tr=1.53 min.

Example 10(2R,5S,11S,14S,17R,18R)-18-Hydroxy-14-isopropyl-2,11,17-trimethyl-15-oxa-3,9,12,28-tetraaza-tricyclo[21.3.1.1*5,9*]octacosa-1(26),23(27),24-triene-4,10,13,16-tetraone—Compound10

To(E)-(2R,5S,11S,14S,17R,18R)-18-(tert-butyl-dimethyl-silanyloxy)-14-isopropyl-2,11,17-trimethyl-15-oxa-3,9,12,28-tetraaza-tricyclo[21.3.1.1*5,9*]octacosa-1(26),21,23(27),24-tetraene-4,10,13,16-tetraone(50 mg, 0.08 mmol) in ethyl acetate (10 mL) at room temperature wasadded 10% palladium on carbon (50 mg). The system was purged withhydrogen gas and stirred vigorously for 16 h. The reaction mixture wasfiltered through Celite and the filter pad washed with methanol. Thefiltrate was concentrated in vacuo. The ensuing residue was dissolved inanhydrous tetrahydrofuran (5 mL) and transferred to a polypropylenevial. At room temperature hydrogen fluoride*pyridine (1.3 mL, 15.2 mmol)was added to the reaction mixture followed by stirring for 3 h. Thereaction was diluted with ethyl acetate and quenched by cautiousaddition of saturated aqueous ammonium chloride. The organic layer wasseparated, dried through a hydrophobic frit and concentrated in vacuo.The residue was purified by silica gel chromatography using ethylacetate/acetone 5/1 to afford the title compound as a white solid. Thiswas further purified by triturating with diethyl ether, filtering anddrying the solid in vacuo (20 mg, 48%, 2 steps). ¹H NMR (300 MHz, CD₃CN)0.90-1.00 (m, 6H), 1.24-1.30 (m, 6H), 1.43 (d, J=6.9 Hz, 3H), 1.32-2.08(m, 11H), 2.45-2.81 (m, 4H), 3.23-3.36 (m, 1H), 3.47-3.59 (m, 1H), 3.67(d, J=10.3 Hz, 1H), 4.02 (d, J=11.8 Hz, 1H), 4.29 (d, J=13.8 Hz, 1H),4.75 (d, J=5.4 Hz, 1H), 4.95-5.08 (m, 1H), 5.16-5.29 (m, 1H), 7.03-7.29(m, 6H). LCMS (m/z)=545.3 [M+H], Tr=2.15 min.

Example 11 Compound 11

Synthesis of Compound 11a:

A cooled (0° C.) solution of (E)-pent-3-en-2-ol (1.024 g, 11.887 mmol)in dichloromethane (30 mL) was subsequently treated with pyridine (1.222g, 1.3 mL, 15.452 mmol) and propionyl chloride (1.430 g, 1.3 mL, 15.452mmol). After stirring for 1.5 h at 0° C. the reaction was quenched withsaturated sodium bicarbonate. The aqueous layer was extracted withdichloromethane and then the organics were combined, filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexane/Et₂O 1:0 to 9:1 toafford the title compound (787 mg, 46%) as a colorless oil.

Synthesis of Compound 11b:

A cooled (0° C.) solution of N,N-diisopropylamine (672.0 mg, 930 μL,6.641 mmol) in tetrahydrofuran (15 mL) was treated with n-butyl lithium(2.5 M in hexanes, 2.4 mL, 6.088 mmol). After stirring at 0° C. for 20min this solution was cooled to −78° C. and subsequently treated withchlorotrimethylsilane (841.7 mg, 990 μL, 7.748 mmol) and a solution of11a (787.0 mg, 5.534 mmol) in tetrahydrofuran (10 mL). The reactionmixture was slowly warmed to room temperature. After 16 h, the reactionwas quenched with 1 M hydrochloric acid (40 mL) and the pH was adjustedto 2 with 2 M hydrochloric acid. The aqueous layer was extracted withethyl acetate (2×40 mL), the organics were combined, dried over sodiumsulfate, filtered and the volatiles were removed in vacuo. The residuewas purified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanelethyl acetate 1:0 to 4:1to afford the title compound (375 mg, 48%) as a colorless oil.

Synthesis of Compound 11c:

Compound 1 ic was prepared in the same manner as Compound 1f using 11band N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride insteadof ic and N,N′-dicyclohexyl carbodiimide in 60% yield.

Synthesis of Compound 11d:

Compound 1d was prepared in the same manner as compound 1k, by firstusing 11c instead of 1f in the preparation of 1g and then, using(R)-1-(3-vinyl-isoquinolin-6-yl)-ethylamine dihydrochloride instead of(R)-1-(3-vinyl-phenyl)-ethylamine hydrochloride following the proceduredescribed for the synthesis of 1k from 1g, in 81% yield.

Synthesis of the Title Compound:

Compound 11 was prepared in the same manner as compound 1 using 11dinstead of 1k and using toluene instead of dichloroethane in 8% yield.¹H NMR (300 MHz, CD₃OD) δ0.97-1.07 (m, 7H), 1.25 (d, J=7.1 Hz, 3H), 1.31(d, J=7.1 Hz, 3H), 1.55-1.63 (m, 6H), 1.65-1.78 (m, 1H), 1.83-2.00 (m,3H), 2.14 (app sextet, J=6.7 Hz, 1H), 2.69-2.79 (m, 1H), 2.80-2.90 (m,1H), 3.65 (app t, J=8.0 Hz, 1H), 4.45 (br d, J=13.6 Hz, 1H), 4.78 (dd,J=6.0, 19.6 Hz, 2H), 5.15 (app pentet, J=6.2 Hz, 1H), 5.83 (dq, J=7.1,19.9 Hz, 1H), 6.65 (dd, J=6.7, 17.2 Hz, 1H), 6.79-6.93 (m, 1H),7.53-7.61 (m, 1H), 7.71 (d, J=5.1 Hz, 1H), 7.96 (d, J=4.7 Hz, 1H), 8.03(d, J=8.0 Hz, 1H), 9.10 (s, 1H). LCMS (m/z) 564.1 [M+H], 596.1 [M+Na],Tr=3.73 min.

Example 12 Compound 12

Synthesis of Compound 12a:

A solution of 2,2-dimethyl-4-pentenoic acid (202 mg, 1.58 mmol) indichloromethane (5 mL) was prepared and oxalyl chloride (411 μL, 4.74mmol) was added followed by dimethylformamide (5 μL). The mixtureeffervesced vigorously and was stirred at room temperature for 1 hbefore evaporating to give a solid (241 mg). The solid was dissolved intoluene (10 mL) and 1e (433 mg, 1.0 mmol) was added followed by silvercyanide (310 mg, 2.25 mmol). The flask was covered with foil to excludelight, before heating to 80° C. for 10 min. It was left at roomtemperature overnight. The reaction was heated to 80° C. for a further4.5 h then allowed to cool. It was filtered and evaporated to give apale yellow gum. The gum was purified by silica gel chromatography usingethyl acetateliso-hexane 1:3 then ethyl acetateliso-hexane 1:1 to yieldthe title compound (200 mg, 37%) as a colorless oil.

Synthesis of Compound 12b:

A solution of 12a (100 mg, 0.184 mmol) in tetrahydrofuran (10 mL) wasprepared and zinc powder (360 mg, 5.52 mmol) was added, followed by pH 6phosphate buffer (1 M, 2 mL). The reaction was stirred at roomtemperature for 18 h and further zinc powder (120 mg, 1.84 mmol) wasadded. The reaction was stirred at room temperature for a further 48 h.The reaction mixture was filtered to remove a suspended solid. The solidwas washed with tetrahydrofuran (15 mL) and pH 6 buffer (5 mL) and thiswas added to the filtrate. The solution was evaporated and the residuepurified by C18 chromatography using a gradient acetonitrile/water 0:1to 1:3 to 1:1. This yielded the title product (69 mg, 91%) as acolorless oil.

Synthesis of Compound 12c:

A solution of 12b (94 mg, 0.23 mmol) in anhydrous acetonitrile (5 mL)was prepared and cooled to 0° C. before adding 2j (54 mg, 0.23 mmol.),N,N-diisopropylethylamine (160 μL, 0.92 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate methanaminium (122 mg, 0.32 mmol). The reaction waswarmed to room temperature and stirred for 16 h. The reaction mixturewas evaporated onto silica gel and purified by silica gel chromatographyusing ethyl acetate to give the title product (119 mg, 87%) as acolorless solid.

Synthesis of the Title Compound:

A solution of 12c (119 mg, 0.20 mmol) in toluene (65 mL) was preparedand Hoveyda-Grubbs 2^(nd) generation catalyst (12.5 mg, 0.02 mmol) wasadded. The reaction mixture was heated to 110° C. under a nitrogenatmosphere for 2 h. A further quantity of Hoveyda-Grubbs 2^(nd)generation catalyst (6 mg, 0.01 mmol) was added and the reaction wasstirred at 110° C. for a further 1 h. The reaction mixture was cooled toroom temperature then filtered, before adsorbing onto silica gel. Thematerial was purified by silica gel chromatography using a gradient ofethyl acetate/acetone 1:0 to 9:1. The resultant gum was triturated withdiethyl ether and dried under vacuum to yield a white solid (29 mg).This solid was further purified by preparatory reverse phase preparativeHPLC to yield the title product (17 mg, 15%) as a white solid. ¹H NMR(300 MHz, CD₃OD) 1.08 (d, J=6.9 Hz, 3H), 1.09 (d, J=6.7 Hz, 3H), 1.18(s, 3H), 1.38 (s, 3H), 1.59 (d, J=3.3 Hz, 3H), 1.61 (d, J=3.1 Hz, 3H),1.83-2.00 (m, 3H), 2.32-2.43 (m, 1H), 2.75-2.82 (m, 2H), 3.60-3.70 (m,1H), 4.40-4.50 (m, 1H), 5.15 (q, J=6.9 Hz, 1H), 6.65 (d, J=16.3 Hz, 1H),7.57 (dd, J=8.5, 1.6 Hz, 1H), 7.70 (s, 1H), 8.00 (s, 1H), 8.03 (d, J=8.7Hz, 1H), 9.10 (s, 1H). LCMS (m/z) 564.1 [M+H], Tr=3.57 min.

Example 13 Compound 13

Synthesis of Compound 13a:

A cooled (0° C.) solution of(S)-4-benzyl-3-{(R)-2-[2-(2-methyl-[1,3]dioxolan-2-yl)-ethyl]-pent-4-enoyl}-oxazolidin-2-one(939.7 mg, 2.516 mmol) as prepared in WO 2001/024797 intetrahydrofuran/water (45 mL, 2:1) was subsequently treated withhydrogen peroxide (30% in water, 1.3 mL, 12.580 mmol) and lithiumhydroxide (211.1 mg, 5.032 mmol). After 2.5 h at 0° C., the reaction wasquenched with sodium hydrogen sulphite (1.4 g). After 0.5 h, thevolatiles were removed in vacuo and the residue diluted with water. Theaqueous layer was extracted with dichloromethane (2×) then acidified topH ˜2 and extracted with dichloromethane (3×). These last organics werecombined, dried over sodium sulfate, filtered and the volatiles wereremoved in vacuo to provide the title compound (538 mg, quant.) as acolorless oil.

Synthesis of Compound 13b:

Compound 13b was prepared in the same manner as compound 1f using 13aand N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride insteadof ic and N,N′-dicyclohexyl carbodiimide in 58% yield.

Synthesis of Compound 13c:

Compound 13c was prepared in the same manner as Compound 1k, by firstusing 13b instead of 1f in the synthesis of 1g, and then using(R)-1-(3-vinyl-isoquinolin-6-yl)-ethylamine dihydrochloride instead of(R)-1-(3-vinyl-phenyl)-ethylamine hydrochloride following the proceduredescribed for the synthesis of 1k from 1g, in 73% yield.

Synthesis of the Title Compound:

Compound 13 was prepared in the same manner as compound 1 using 13cinstead of 1k and using toluene instead of dichloroethane in 32% yield.¹H NMR (300 MHz, CD₃OD) δ0.88-1.00 (m, 1H), 1.06 (2d, J=6.9 Hz, 6H),1.22-1.42 (m, 4H), 1.52-1.63 (m, 7H), 1.65-1.81 (m, 3H), 1.84-1.99 (m,3H), 2.16 (app pentet, J=6.9 Hz, 1H), 2.38-2.50 (m, 1H), 2.51-2.61 (m,1H), 2.62-2.74 (m, 1H), 2.83 (dt, J=2.7, 13.8 Hz, 1H), 3.59-3.72 (m,1H), 4.46 (br dd, J=3.3, 13.6 Hz, 1H), 4.69-4.79 (m, 4H), 5.16 (q, J=7.1Hz, 1H), 5.89 (q, J=6.9 Hz, 1H), 6.71 (d, J=16.3 Hz, 1H), 6.84-6.98 (m,1H), 7.59 (d, J=8.5 Hz, 1H), 7.71 (s, 1H), 7.96 (s, 1H), 8.03 (d, J=8.7Hz, 1H), 9.10 (s, 1H). LCMS (m/z) 651.2 [M+H], Tr=3.68 min.

Example 14 Compound 14

A solution of 13 (183.7 mg, 0.282 mmol) in acetone (10 mL) was treatedwith para-toluene sulfonic acid (27 mg, 0.141 mmol). After stirring atroom temperature for 2.5 h, the reaction was quenched with a saturatedsolution of sodium bicarbonate and the aqueous layer was extracted withethyl acetate (2×20 mL). Organics were combined, dried over sodiumsulfate, filtered and the volatiles were removed in vacuo. The residuewas purified by silica gel chromatography using a 25 g Isolute cartridgeeluted with a continuous gradient of iso-hexane/acetone 1:0 to 2:3 andthen triturated with iso-hexanelacetone to provide the title compound(49.3 mg, 28%) as a white solid. ¹H NMR (300 MHz, CD₃OD) 1.04 (d, J=6.9Hz, 3H), 1.05 (d, J=6.9 Hz, 3H), 1.53-1.62 (m, 7H), 1.64-1.79 (m, 1H),1.84-2.03 (m, 5H), 2.10-2.23 (m, 4H), 2.39-2.53 (m, 1H), 2.54-2.73 (m,4H), 2.82 (dt, J=2.9, 13.1 Hz, 1H), 3.59-3.70 (m, 1H), 4.46 (br dd,J=4.0, 12.7 Hz, 1H), 5.17 (q, J=6.9 Hz, 1H), 5.91 (, J=7.1 Hz, 1H), 6.71(d, J=16.3 Hz, 1H), 6.86-7.00 (m, 1H), 7.59 (d, J=8.7 Hz, 1H), 7.71 (s,1H), 7.98 (s, 1H), 8.03 (d, J=8.7 Hz, 1H), 9.10 (s, 1H). LCMS (m/z)606.1 [M+H], Tr=3.26 min.

Example 15 Compound 15

A stock solution of O-(2-morpholin-4-yl-ethyl)-hydroxylamine (18 mg,0.125 mmol) and acetic acid (a drop) in 2 mL isopropanol was prepared.To solid Compound 14 (16.4 mg, 0.027 mmol) was added 1 mL of the stocksolution and after stirring at room temperature for 18 h the compoundwas purified by reverse phase HPLC to provide a mixture of geometricisomers of the oxime (1:1, 13.3 mg, 67%) as a white solid. ¹H NMR (300MHz, CD₃OD) 1.00-1.10 (m, 6H), 1.54-1.64 (m, 5H), 1.65-1.78 (m, 1H),1.81-2.04 (m, 8H), 2.09-2.22 (m, 1H), 2.23-2.33 (m, 1H), 2.34-2.90 (m,12H), 3.53-3.76 (m, 6H), 4.18 (dt, J=5.8, 8.5 Hz, 2H), 4.41-4.51 (m,1H), 4.68-4.82 (m, 2H), 5.16 (q, J=7.3 Hz, 1H), 5.82-5.96 (m, 1H), 6.71(d, J=15.8 Hz, 1H), 6.84-7.00 (m, 1H), 7.59 (d, J=8.7 Hz, 1H), 7.71 (s,1H), 7.98 (d, J=2.9 Hz, 1H), 8.04 (d, J=8.7 Hz, 1H), 8.53 (br dd, J=3.1,9.4 Hz, 1H), 9.10 (s, 1H). LCMS (m/z) 734.7 [M+H], Tr=1.08 min.

Example 16 Compound 16

A solution of sodium triacetoxyborohydride (203.5 mg, 0.96 mmol) inmethanol (1 mL) and acetic acid (100 μL) was subsequently treated with a2 M solution of dimethylamine in methanol (480 μL, 0.96 mmol) andcompound 14 (5.8 mg, 0.0096 mmol). After stirring for a month at roomtemperature, the reaction mixture was directly loaded onto the reversephase preparative HPLC which was eluted with a gradient of water/CH₃CN95:5 to 0:100. This provided the title compound (1.1 mg, 18%) as a whitesolid and as a 1:1 mixture of diastereomers. ¹H NMR (300 MHz, CD₃OD)δ0.87-0.97 (m, 1H), 1.01-1.09 (m, 6H), 1.24 (d, J=6.4 Hz, 3H), 1.55-1.63(m, 6H), 1.64-1.80 (m, 2H), 1.84-2.02 (m, 4H), 2.10-2.24 (m, 1H),2.41-2.72 (m, 7H), 2.73-2.90 (m, 3H), 3.60-3.69 (m, 1H), 4.41-4.51 (m,1H), 5.17 (q, J=7.1 Hz, 1H), 5.85-5.95 (m, 1H), 6.71 (d, J=16.0 Hz, 1H),6.85-6.99 (m, 1H), 7.56-7.63 (m, 1H), 7.71 (s, 1H), 7.98 (s, 1H), 8.04(d, J=8.5 Hz, 1H), 9.06-9.13 (m, 1H). LCMS (m/z) 635.0 [M+H], Tr=0.94min.

Example 17(E)-(2R,5S,11S,14S)-14-Isopropyl-2,11,15-trimethyl-3,9,12,15,28-pentaaza-tricyclo[21.3.1.1*5,9*]octacosa-1(27),21,23,25-tetraene-4,10,13,16-tetraone—Compound17

Synthesis of Compound 17a:

A solution of 1d (1.08 g, 2.5 mmol) in dichloromethane (35 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (682 mg, 0.55 mL, 3.1 mmol) was added and thereaction mixture was stirred at 0° C. for 45 minutes.N,N-diisopropylethylamine (1.29 g, 1.73 mL, 10 mmol) was added and thesolvent was evaporated to afford crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (2.5 mmol), which was used in the next step.A solution of crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (2.5 mmol) in acetonitrile (30 mL) wasstirred at 0° C. under nitrogen.(S)-2-(tert-Butoxycarbonyl-methyl-amino)-3-methyl-butyric acid (635 mg,2.75 mmol) and N,N-diisopropylethylamine (1.29 g, 1.7 mL, 10 mmol) wasadded followed by 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate methanaminium (1.33 g, 3.5 mmol) and thereaction mixture was stirred at room temperature for 18 h. The solventwas evaporated and the residue was dissolved in ethyl acetate. Thesolution was washed with water, 2 M hydrochloric acid, water, saturatedsodium hydrogen carbonate solution, water and brine, and then dried overanhydrous sodium sulfate, filtered and evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanelethyl acetate 1:1 to neat ethyl acetate to afford the titlecompound (950 mg, 70%) as a white solid.

Synthesis of Compound 17b:

A solution of 17a (950 mg, 1.74 mmol) in dichloromethane (25 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (577 mg, 0.47 mL, 2.6 mmol) was added and thereaction mixture was stirred at 0° C. for 1 h. Additional trimethylsilyltrifluoromethanesulfonate (0.2 mL, 1.1 mmol) was added and the reactionmixture was stirred at 0° C. for 2 h. N,N-diisopropylethylamine (903 mg,1.2 mL, 7 mmol) was added and the solvent was evaporated and the residuewas dried to afford crude(S)-1-[(S)-2-((S)-3-methyl-2-methylamino-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (1.74 mmol), which was used in the nextstep. A solution of crude(S)-1-[(S)-2-((S)-3-methyl-2-methylamino-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (1.74 mmol) in acetonitrile (20 mL) wasstirred at 0° C. under nitrogen. Hept-6-enoic acid (268 mg, 0.3 mL, 2.1mmol) and N,N-diisopropylethylamine (903 mg, 1.2 mL, 7 mmol) were added.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (912 mg, 2.4 mmol) was added and thereaction mixture was stirred at room temperature for 18 h. The solventwas evaporated and the residue was diluted with ethyl acetate. Thesolution was washed with saturated sodium hydrogen carbonate solution,water, 2 M hydrochloric acid, water and brine, dried over anhydroussodium sulfate, filtered and evaporated. The residue was purified bysilica gel chromatography using a gradient of iso-hexanelethyl acetate3:1 to 1:1 to afford the title compound (710 mg, 74%) as a white solid.

Synthesis of Compound 17c:

A solution of 17b (710 mg, 1.3 mmol) in tetrahydrofuran (30 mL) wasstirred at room temperature under nitrogen. Zinc dust (1.86 g, 28.6mmol) was added followed by a solution of ammonium acetate (1.50 g, 19.5mmol) in water (20 mL) and the reaction mixture was stirred at roomtemperature for 24 hours. The reaction mixture was filtered throughCelite and the filter pad was washed with ethyl acetate and potassiumhydrogen sulfate solution (pH ˜2). The filtrate was acidified to pH 2with 2 M hydrochloric acid and the mixture was extracted with ethylacetate. The organic extracts were combined, washed with water andbrine, dried over anhydrous sodium sulfate, filtered and evaporated. Theresidue was co-evaporated with toluene and dried to afford(S)-1-{(S)-2-[(S)-2-(hept-6-enoyl-methyl-amino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (468 mg, 85%) as a gum. A solution of(S)-1-{(S)-2-[(S)-2-(hept-6-enoyl-methyl-amino)-3-methyl-butyrylamino]propionyl}-hexahydropyridazine-3-carboxylicacid (468 mg, 1.1 mmol) in acetonitrile (25 mL) was stirred at roomtemperature under nitrogen. 1j (200 mg, 1.1 mmol) andN,N-diisopropylethylamine (710 mg, 1.0 mL, 5.5 mmol) was added followedby 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (585 mg, 1.54 mmol) and the reactionmixture was stirred at room temperature for 72 hours. The solvent wasevaporated and the residue was dissolved in ethyl acetate. The solutionwas washed with 2 M hydrochloric acid, water, saturated sodium hydrogencarbonate solution, water and brine, dried over anhydrous sodiumsulfate, filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanelethyl acetate 1:1 to ethylacetate to afford the title compound as a white solid (300 mg, 49%).

Synthesis of the Title Compound:

A solution of 17c (276 mg, 0.5 mmol) in 1,2-dichloroethane (150 mL) wasstirred at room temperature under nitrogen. Hoveyda-Grubbs 2ndgeneration catalyst (32 mg, 0.05 mmol) was added and the reactionmixture was heated at 80° C. under nitrogen for 3 hours. The reactionmixture was cooled to room temperature, silica gel was added and thereaction mixture was evaporated. The residue was purified by silica gelchromatography using a gradient of ethyl acetate to ethylacetate/acetone 7:3. The residue was triturated with ether and theresulting solid was collected, washed with ether and dried to afford thetitle compound (112 mg, 43%) as a white solid. ¹H NMR (300 MHz, CD₃CN)60.82 (d, J=6.7 Hz, 3H), 0.90 (d, J=6.5 Hz, 3H), 1.34 (d, J=7.1 Hz, 3H),1.39 (d, J=6.3 Hz, 3H), 1.40-1.90 (m, 10H), 2.60-2.75 (m, 2H), 2.86-2.92(m, 4H), 3.28-3.36 (m, 1H), 3.93 (d, J=12.0 Hz, 1H), 4.33-4.38 (m, 1H),4.58 (d, J=11.6 Hz, 1H), 4.85-4.99 (m, 2H), 6.16-6.51 (m, 3H), 7.12-7.30(m, 6H). LCMS (m/z) 526.3 [M+H], Tr=2.16 min.

Example 18 Compound 18

Synthesis of Compound 18a:

To a stirred slurry of 7-bromo-2-chloro-quinoline (Fluorochem, 8.10 g,33.4 mmol) and sodium iodide (50.0 g, 334 mmol) in acetonitrile (27 mL)was added acetyl chloride (3.56 mL, 50.0 mmol) slowly. The flask wasstoppered, sealed and heated at 80° C. for 3 h before being allowed tocool. The mixture was treated sequentially with 10% w/w aqueouspotassium carbonate solution (80 mL), 5% w/w aqueous sodium sulfitesolution (80 mL) and saturated aqueous sodium thosulfate solution (80mL) and the mixture extracted with dichloromethane (2×). The combinedorganic extracts were dried over sodium sulfate, filtered and evaporatedto give a crude 7-bromo-2-iodo-quinoline. To the quinoline was addedtributyl(1-ethoxyvinyl)stannane (13.6 mL, 40.1 mmol), 1,4-dioxane (67mL) and bis(triphenylphosphine)palladium(II) dichloride (2.37 g, 3.34mmol) and the reaction mixture heated at 100° C. for 5 h before beingallowed to cool. 2 M aqueous hydrochloric acid (67 mL) was added and thereaction stirred for 1 h. The mixture was filtered and the solids washedwith ethyl acetate and the filtrate evaporated to remove organics. Theresidue was extracted with ethyl acetate (3×) and the combined organicextracts were dried over sodium sulfate, filtered and evaporated. Theproduct was purified on silica gel doped with 10% w/w potassiumcarbonate, eluting with a gradient of 0 to 6% ethyl acetate iniso-hexanes to afford the title compound (5.5 g, 66%) as a white solid.

Synthesis of Compound 18b:

To a solution of 18a (1.42 g, 5.68 mmol) in tetrahydrofuran (28 mL) wasadded titanium (IV) ethoxide (2.6 g, 2.35 mL, 11.4 mmol) followed by(R)-(+)-2-methyl-propanesulfinimide (825 mg, 6.82 mmol). The reactionmixture was stirred at 60° C. under nitrogen for 6 h and allowed tocool. Brine was added followed by ethyl acetate and the suspensionfiltered through celite and the filter pad was washed with ethylacetate. The ethyl acetate layer was separated, dried over sodiumsulfate, filtered and evaporated. The residue was purified by silica gelchromatography eluting with 10-25% ethyl acetate in iso-hexanes toafford the title compound (448 mg, 22%).

Synthesis of Compound 18c:

A mixture of (1S,2R)-(−)-cis-1-amino-2-indanol (19 mg, 0.13 mmol),[Ru(p-cymen)Cl₂]₂ (39 mg, 0.064 mmol) and powdered 4 Å molecular sieves(0.7 g) was suspended in anhydrous 2-propanol (3 mL) and stirred undernitrogen. The suspension was heated at 90° C. for 30 min. The reactionmixture was cooled to 40° C. and a solution of 18b (448 mg, 1.27 mmol)in 2-propanol (9 mL) was added followed by a solution of potassiumtert-butoxide (36 mg, 0.32 mmol) in 2-propanol (3 mL). The reactionmixture was stirred for 2 h at 40° C. and then allowed to cool. Themixture was poured directly onto a silica gel cartridge and eluted withethyl acetate. After concentration the residue was further purified onsilica eluting with ethyl acetate in iso-hexanes (1:1 to 1:0) to affordthe title compound (287 mg, 64%).

Compound 18d:

Synthesis of Compound 18d: A mixture of 18c (207 mg, 0.583 mmol),(E)-2,2-Dimethyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-but-3-enoicacid methyl ester (170 mg, 0.670 mmol),bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)chloride (24 mg, 0.029 mmol) and potassium phosphate tribasic (371 mg,1.75 mmol) was suspended in cyclopentylmethyl ether (3 mL) and water(1.5 mL) and the mixture stirred and heated at 80° C. under nitrogen for5 h. The reaction mixture was allowed to cool and diluted with ethylacetate and water. The aqueous layer was separated and washed withfurther ethyl acetate and the combined organic extracts dried oversodium sulfate, filtered and evaporated. The residue was purified bysilica gel chromatography using a gradient of ethyl acetate iniso-hexanes (1:1 to 3:1) to afford the title compound as a yellow oil(201 mg, 86%).

Synthesis of Compound 18e:

18d (193 mg, 0.48 mmol) was suspended in 4 M HCl in 1,4-dioxane (5 mL,20 mmol) and methanol was added (10 mL). The reaction mixture wasstirred for 2 h and then evaporated. The residue was triturated withether to give the title compound as a yellow solid (166 mg).

Synthesis of Compound 18f:

(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (166 mg, 0.48 mmol), 18e (190 mg, 0.48 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (256 mg, 0.67 mmol) and powdered 4 Å molecularsieves were suspended in acetonitrile (9.6 mL) under nitrogen. To thestirred mixture was added N,N-diisopropylethylamine (186 mg, 250 μL,1.44 mmol) and the reaction mixture stirred for 20 h. The mixture wasfiltered through a phase separator and the filtrate evaporated,dissolved in dichloromethane and washed successively with saturatedsodium bicarbonate solution, water and saturated ammonium chloridesolution, and then dried over sodium sulfate, filtered and evaporated.The residue was purified by silica gel chromatography using a gradientof ethyl acetate in iso-hexanes (1:1 to 1:0) to afford the titlecompound as a yellow gum (77 mg, 28% over 2 steps).

Synthesis of Compound 18g:

To 18f (117 mg, 0.201 mmol) in tetrahydrofuran (4 mL) was added, withstirring, water (0.8 mL) and lithium hydroxide monohydrate (9.2 mg,0.221 mmol). The reaction mixture was stirred for 1 h and then furtherlithium hydroxide monohydrate (36 mg, 0.857 mmol) was added and themixture stirred for a further 19.5 hours. The reaction mixture wasevaporated and the residue azeotroped with toluene. The residue waspurified by preparative HPLC using a gradient of 20 to 100% acetonitrilein water to give 18g as a white solid (60 mg, 53%).

Synthesis of the Title Compound:

A slurry of 2-methyl-6-nitrobenzoic anhydride (177 mg, 0.500 mmol), DMAP(92 mg, 0.750 mmol) and powdered 4 Å molecular sieves (1 g) in1,2-dichloroethane (33 mL) was stirred under nitrogen at 50° C. 18g (57mg, 0.100 mmol) was dissolved in N,N-dimethylformamide (5 mL) and thesolution added to the slurry over 5 h via syringe pump. After theaddition was complete the original flask containing the acid was washedwith further N,N-dimethylformamide (1 mL) and the washings transferredto the reaction vessel over 30 min. The reaction mixture was stirred at50° C. for a further hour and then allowed to cool. The residue waspurified by preparative HPLC using a gradient of 5 to 100% acetonitrilein water to give the title compound as a white solid (24.5 mg, 45%). ¹HNMR (300 MHz, CD₃OD) δ 1.00 (d, J=6.7 Hz, 3H), 1.07 (d, J=6.7 Hz, 3H),1.43 (s, 3H), 1.54 (s, 3H), 1.59 (d, J=7.6 Hz, 3H), 1.64 (d, J=7.4 Hz,3H), 1.96 (m, 1H), 2.17 (m, 1H), 2.28 (m, 1H), 2.71 (m, 1H), 3.61 (m,1H), 4.43 (m, 1H), 5.10 (m, 1H), 5.24 (d, J=8.9 Hz, 1H), 5.81 (q, J=7.1Hz, 1H), 6.46 (ABq, Δδ_(AB)=0.18, J_(AB)=16.3 Hz, 2H), 7.45 (d, J=8.5Hz, 1H), 7.62 (dd, J=8.5, 1.3 Hz, 1H), 7.79 (s, 1H), 7.85 (d, J=8.5 Hz,1H), 8.26 (d, J=8.5 Hz, 1H), 9.41 (br d, J=5.6 Hz, 1H). LCMS (m/z) 550.2[M+H], Tr 2.05 min.

Example 19 Compound 19

Synthesis of Compound 19a:

1e (1.08 g, 2.50 mmol), 2-methyl-6-nitrobenzoic anhydride (1.46 g, 4.25mmol), 4-dimethylaminopyridine (122 mg, 1.00 mmol) and powdered 4 Åmolecular sieves were suspended in dry dichloromethane (11 mL) and themixture stirred under nitrogen. Triethylamine (633 mg, 871 μL, 6.25mmol) and a solution of 2,2-dimethyl-but-3-enoic acid (390 mg, 3.43mmol) in dry dichloromethane (1.5 mL) were added and the reactionmixture stirred for 17 h. The suspension was passed through a phaseseparator cartridge and the filtrate evaporated. The residue waspurified by silica gel chromatography using a gradient of ethyl acetatein iso-hexanes (1:4 to 2:3) to afford the title compound as a brown gum(735 mg, 56%).

Synthesis of Compound 19b:

19a (124 mg, 0.235 mmol), (R)-1-(7-bromo-quinolin-2-yl)-ethanol (65 mg,0.258 mmol), palladium acetate (8 mg, 0.035 mmol), tri(o-tolyl)phosphine(22 mg, 0.071 mmol) and triethylamine (48 mg, 66 μL, 0.47 mmol) weresuspended in acetonitrile (2.35 mL). The reaction mixture was heated andstirred at 120° C. for 2 h in a microwave reactor. The residue wasevaporated and suspended in ethyl acetate and saturated ammoniumchloride solution. The organic layer was separated and washed withsaturated ammonium chloride solution then water and the combined aqueouswashes back-extracted with ethyl acetate. The combined organic layerswere washed with brine, dried over sodium sulfate, filtered andevaporated. The residue was purified by silica gel chromatography usinga gradient of ethyl acetate in iso-hexanes (1:1 to 3:1) to afford thetitle compound as a yellow gum (38 mg, 23%).

Synthesis of Compound 19c:

To a solution of 19b (58 mg, 0.083 mg) in tetrahydrofuran (5 mL) wasadded zinc (218 mg, 3.33 mmol) and pH 6 phosphate buffer (1 mL). Thereaction mixture was rapidly stirred for 18 h. Zinc (218 mg, 3.33 mol)was added and the reaction heated to 40° C. for 1 h and allowed to cool.The mixture was filtered through a phase separator washing withtetrahydrofuran and pH 6 phosphate buffer. The filtrate was diluted withethyl acetate and water and the aqueous phase extracted with furtherethyl acetate. The combined organic extracts were dried over sodiumsulfate, filtered and evaporated. The aqueous phase was evaporated toca. half volume and extracted with ethyl acetate (5×) and the combinedorganic extracts dried over sodium sulfate, filtered and combined withthe rest of the product and evaporated to give a yellow gum (51 mg).

Synthesis of the Title Compound:

A slurry of 2-methyl-6-nitrobenzoic anhydride (143 mg, 0.415 mmol),4-dimethylaminopyridine (76 mg, 0.623 mmol) and powdered 4 Å molecularsieves (1 g) in 1,2-dichloroethane (27 mL) was stirred under nitrogen at50° C. 19c (51 mg, 0.083 mmol) was dissolved in N,N-dimethylformamide (3mL) and the solution added to the slurry over 3.5 h via syringe pump.After the addition was complete the original flask containing the acidwas washed with further N,N-dimethylformamide (1 mL) and the washingstransferred to the reaction vessel over 10 min. The reaction mixture wasstirred at 50° C. for a further hour and then allowed to cool. Theresidue was purified by preparative HPLC using a gradient of 10% to 100%acetonitrile in water. The residue was further purified by silica gelchromatography using a gradient of 50% to 60% ethyl acetate iniso-hexanes (1:1 to 3:2) to give the title compound as a colorless gum(1.7 mg, 4% over 2 steps). ¹H NMR (300 MHz, CD₃OD) δ 0.94 (d, J=7.0 Hz,3H), 0.99 (d, J=6.7 Hz, 3H), 1.41 (s, 3H), 1.52 (s, 3H), 1.65 (d, J=7.1Hz, 3H), 1.68 (m, 2H), 1.73 (d, J=6.7 Hz, 3H), 1.89-2.03 (m, 2H), 2.15(m, 1H), 2.76 (m, 1H), 3.82 (m, 1H), 4.39 (br d, J=13.4 Hz, 1H), 5.08(d, J=9.8 Hz, 1H), 5.81 (q, J=7.1 Hz, 1H), 5.93 (q, J=6.9 Hz, 1H), 6.34(ABq, Δδ_(AB)=0.11, J_(AB)=16.6 Hz, 2H), 7.41 (d, J=8.5 Hz, 1H), 7.67(s, 1H), 7.75 (dd, J=8.7, 1.3 Hz, 1H), 7.82 (d, J=8.7 Hz, 1H), 8.22 (d,J=8.5 Hz, 1H). LCMS (m/z) 551.2 [M+H], Tr 2.22 min.

Example 20 Compound 20

Synthesis of Compound 20a:

Dichloro(p-cumene)ruthenium(II) dimer (3 mg, 0.005 mmol) and(1R,2R)-(−)—N-p-tosyl-1,2-diphenylethylenediamine (4.4 mg, 0.012 mmol)were suspended in degassed water (2 mL) and the mixture was degassedwith nitrogen for 15 min. The mixture was stirred at 70° C. undernitrogen for 90 min. The resulting yellow solution was cooled to roomtemperature. 2f (206 mg, 1 mmol), sodium formate (340 mg, 5 mmol) anddegassed tetrahydrofuran (1 mL) were added and the reaction mixture wasdegassed with nitrogen for 5 min. The reaction mixture was vigorouslystirred at 40° C. for 2.5 h. The reaction mixture was cooled to roomtemperature and was extracted with ethyl acetate. The organic layer wasseparated, washed with water and brine, dried over sodium sulfate,filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 4:1 to 2:1to afford the title compound (193 mg, 92%) as a white solid.

Synthesis of Compound 20b:

1,4-Dioxane (5 mL) was degassed with nitrogen, 20a (208 mg, 1 mmol),tributyl(vinyl)tin (951 mg, 0.9 mL, 3 mmol) andbis(triphenylphosphine)palladium(II) dichloride (70 mg, 0.1 mmol) wereadded and the reaction mixture was heated at 150° C. in a microwavereactor for 1 h. Additional tributyl(vinyl)tin (0.3 mL, 1 mmol) andbis(triphenylphosphine)palladium(II) dichloride (70 mg, 0.1 mmol) wereadded and the reaction mixture was heated at 150° C. in a microwavereactor for 1 h. The reaction mixture was cooled to room temperature andthe mixture was filtered through filter aid and the filter pad waswashed with ethyl acetate. The filtrate was evaporated and the residuewas purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 4:1 to 2:1 followed by silica gelchromatography using iso-hexanes/ethyl acetate 3:1 to afford the titlecompound (100 mg, 50%) as a white solid.

Synthesis of Compound 20c:

A solution of 1d (6.88 g, 15.9 mmol) in dichloromethane (200 mL) wasprepared and trifluoroacetic acid (50 mL) was added. The reactionmixture was stirred at room temperature for 2 h. TLC showed the reactionto be complete. The solution was evaporated to give a brown oil. Thiswas azeotroped with toluene (50 mL) and the resultant oil was driedunder vacuum to give(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester trifluoroacetic acid salt (7.8 g) as a browngum. A solution of ((S)-1-carbamoyl-2-methylpropyl)-carbamic acidtert-butyl ester in acetonitrile (300 mL) was cooled to 0° C. beforeadding N,N-diisopropylethylamine (13.8 mL, 79.7 mmol) and2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(6.33 g, 16.7 mmol). The reaction was stirred at 0° C. for 15 min beforeadding a solution of the(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester trifluoroacetic acid salt (ca. 15.9 mmol) inacetonitrile (85 mL). The reaction was stirred at 0° C. for a further 20min and then allowed to warm to room temperature and stirred for 15 h.The reaction mixture was evaporated then re-dissolved in ethyl acetate(250 mL). The solution was washed with water (150 mL) then dried oversodium sulfate, filtered and evaporated to give a red oil. This waspurified by silica gel chromatography using iso-hexanelethyl acetate 7:3then iso-hexanelethyl acetate 1:1 to yield the title compound (8.2 g,92%) as a pale orange amorphous solid.

Synthesis of Compound 20d:

To a solution of 20c (530 mg, 1.00 mmol) in tetrahydrofuran (3 mL) andwater (2 mL) was added lithium hydroxide (26 mg, 1.1 mmol) at 23° C.under an argon atmosphere. After 19 h, the reaction mixture wasconcentrated under reduced pressure and the crude residue was purifiedby silica gel flash column chromatography (12 g Combiflash HP GoldColumn, 0-20% methanol/dichloromethane gradient) to afford the titlecompound (339 mg, 92%) as a colorless solid.

Synthesis of Compound 20e:

To a solution of 20d (339 mg, 0.920 mmol) and 20 bl (183 mg, 0.920 mmol)in dichloromethane (4.6 mL) were addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (211 mg,1.10 mmol) and 4-dimethylaminopyridine (56.2 mg, 0.46 mmol) at 23° C.under an argon atmosphere. After 21 h, the reaction mixture was purifieddirectly by silica gel flash column chromatography (24 g Combiflash HPGold Column, 0-100% ethyl acetate/hexanes gradient) to afford the titlecompound (224 mg, 42%) as a light tan solid.

Synthesis of Compound 20f:

To a solution of 20e (224 mg, 0.390 mmol) in dichloromethane (2.0 mL)was added trimethylsilyl trifluoromethanesulfonate (128 mg, 0.58 mmol)at 0° C. under an argon atmosphere. After 2 h, the reaction mixture wasconcentrated under reduced pressure to afford the title compound (188mg, 99%) as a light yellow oil, which was used without furtherpurification.

Synthesis of Compound 20g:

To a solution of 20f (94 mg, 0.195 mmol) in dimethylformamide (1.00 mL)were added cesium hydroxide monohydrate (98.5 mg, 0.585 mmol) andactivated 4 Å molecular sieves (53 mg) at 23° C. under an argonatmosphere. After 10 min, 5-bromopent-1-ene (29 mg, 0.195 mmol) wasadded. After 4 d, the reaction mixture was diluted with brine (20 mL)and was extracted with ethyl acetate (3×20 mL). The combined organicextracts were dried over anhydrous sodium sulfate and were concentratedunder reduced pressure. The crude residue was purified by silica gelflash column chromatography (12 g Combiflash HP Gold Column, 0-20%methanol/dichloromethane gradient) to afford the title compound (5.7 mg,5%) as a light yellow solid.

Synthesis of the Title Compound:

To a solution of 20g (5.7 mg, 11 μmol) in toluene (3.7 mL) was added theHoveyda-Grubbs 2nd Generation Catalyst (1.0 mg, 1.7 μmol) at 23° C.under an argon atmosphere, and the resulting mixture was heated to 110°C. After 1.25 h, the reaction mixture was allowed to cool to 23° C., andwas concentrated under reduced pressure. The crude residue was purifiedby preparatory HPLC (Gemini 5 u C18 110 Å column, 5-100% MeCN/H₂O, 0.1%trifluoroacetic acid modifier) to afford the title compound (3.7 mg,56%) as a white solid trifluoroacetic acid salt. ¹H NMR (CD₃OD, 400MHz): δ9.21 (s, 1H), 8.09 (d, J=8.6 Hz, 1H), 7.84 (s, 1H), 7.79 (s, 1H),7.60 (d, J=8.6 Hz, 1H), 6.66 (d, J=15.9 Hz, 1H), 6.57-6.45 (m, 1H), 5.99(q, J=6.7 Hz, 1H), 5.30 (q, J=7.0 Hz, 1H), 4.31 (br-d, J=12.0 Hz, 1H),3.69-3.61 (m, 2H), 2.91 (br-t, J=8.1 Hz, 2H), 2.68 (td, J=12.8, 3.2 Hz,1H), 2.55-2.44 (m, 1H), 2.42-2.31 (m, 1H), 2.15-2.03 (m, 1H), 1.99-1.78(m, 4H), 1.74-1.59 (m, 2H), 1.60 (d, J=6.7 Hz, 3H), 1.56 (d, J=7.1 Hz,3H), 1.06 (d, J=6.9 Hz, 3H), 0.99 (d, J=6.9 Hz, 3H). HPLC t_(R) (min),4.169, purity %: 99%. (Phenomenex Kinetex 2.6 u C18 100 Å, 100×4.60 mm2.6 micron column, 8.5 min, 1.5 ml/min, 2-98% MeCN/H₂O, 0.1%trifluoroacetic acid modifier gradient). LCMS (ESI) m/z 522.2 [M+H]⁺,t_(R)=1.68 min. (Gemini 5 u C18 110 Å, 50×4.60 mm 5 micron column, 3.5min, 2 ml/min, 5-100% MeCN/H₂O, 0.1% acetic acid modifier gradient).

Example 21 Compound 21

Synthesis of Compound 21a:

To a solution of 20c (500 mg, 0.945 mmol) in dichloromethane (4.7 mL)was added trimethylsilyl trifluoromethanesulfonate (314 mg, 0.1.41 mmol)at 0° C. under an argon atmosphere. After 1 h, the reaction mixture wasconcentrated under reduced pressure. The resulting residue was dilutedwith dichloromethane (4.7 mL) and N,N-diisopropylethylamine (657 μL,3.78 mmol) and prop-2-ene-1-sulfonyl chloride (140 mg, 0.992 mmol) weresequentially added at 23° C. under an argon atmosphere. After 17 h, thereaction mixture was diluted with dichloromethane (50 mL) and theresulting mixture was washed with saturated aqueous sodium bicarbonatesolution (50 mL). The organic layer was separated, dried over anhydroussodium sulfate, and was concentrated under reduced pressure. The cruderesidue was purified by silica gel flash column chromatography (24 gCombiflash HP Gold Column, 0-100% ethyl acetate/hexanes gradient) toafford the title compound (256.4 mg, 51%) as an orange oil.

Synthesis of Compound 21b:

To a solution of 21a (256 mg, 0.478 mmol) in tetrahydrofuran (8.0 mL)were added zinc powder (625 mg, 9.57 mmol) followed by a solution ofammonium acetate (552 mg, 7.17 mmol) in water (5.31 mL) at 23° C. underan argon atmosphere. After 17 h, the reaction mixture was warmed to 45°C. After 7 h, the reaction mixture was allowed to cool to roomtemperature and was filtered through a pad of celite washing with water(10 mL). The filtrate was concentrated under reduced pressure to removetetrahydrofuran and the residue was partitioned between water (25 mL)and ethyl acetate (25 mL). The layers were split and the aqueous layerwas acidified to pH=2 with 12 N aqueous hydrogen chloride solution. Theaqueous layer was extracted with ethyl acetate (4×50 mL), and thecombined organic extracts were dried over anhydrous sodium sulfate, andwere concentrated under reduced pressure. Residual acetic acid wasremoved azeotropically via addition of toluene (5 mL) followed byconcentration under reduced pressure (3×) to afford the title compound(156 mg, 80%) as a white solid.

Synthesis of Compound 21c:

To a solution of 21b (156 mg, 0.390 mmol) and 20b (92.3 mg, 0.464 mmol)in dichloromethane (1.95 mL) were addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (105 mg,0.546 mmol) and 4-dimethylaminopyridine (23.8 mg, 195 μmol) at 23° C.under an argon atmosphere. After 21 h, the reaction mixture was purifieddirectly by silica gel flash column chromatography (24 g Combiflash HPGold Column, 0-100% ethyl acetate/hexanes gradient) to afford the titlecompound (155 mg, 70%) as a colorless oil.

Synthesis of the Title Compound:

To a solution of 21c (150 mg, 256 μmol) in toluene (51 mL) was added theHoveyda-Grubbs 2nd Generation Catalyst (16 mg, 25 μmol) at 23° C. underan argon atmosphere, and the resulting mixture was heated to 110° C.After 2 h, the reaction mixture was quenched with ethyl vinyl ether (500μL) and the resulting mixture was allowed to cool to 23° C. The reactionmixture was concentrated under reduced pressure and the crude residuewas purified by silica gel flash column chromatography (12 g CombiflashHP Gold Column, 0-100% ethyl acetate/hexanes gradient) to afford thetitle compound (18.6 mg, 13%) as a tan solid. ¹H NMR (CD₃OD, 400 MHz):δ9.37 (s, 1H), 8.23 (d, J=8.5 Hz, 1H), 8.14 (s, 1H), 7.97 (s, 1H), 7.77(d, J=8.7 Hz, 1H), 6.85 (d, J=15.8 Hz, 1H), 6.75-6.64 (m, 1H), 6.22 (q,J=6.7 Hz, 1H), 5.62-5.52 (m, 1H), 4.40 (dd, J=13.3, 3.3 Hz, 1H), 4.21(dd, J=14.8, 5.8 Hz, 1H), 4.10 (dd, J=13.4, 7.5 Hz, 1H), 4.06 (d, J=4.5Hz, 1H), 3.72 (dd, J=11.5, 2.5 Hz, 1H), 2.79 (td, J=12.8, 3.2 Hz, 1H),2.19 (app-sextet, J=7.0 Hz, 1H), 2.04-1.88 (m, 2H), 1.87-1.66 (m, 2H),1.72 (d, J=6.7 Hz, 3H), 1.46 (d, J=7.2 Hz, 3H), 1.05 (d, J=6.7 Hz, 3H),0.92 (d, J=6.8 Hz, 3H). HPLC t_(R) (min), 2.882, purity %: 98%. (Synergi4 u hydro-RP, 50×4.60 mm 4 micron column, 7 min, 2 ml/min, 5-100%MeCN/H₂O, 0.05% trifluoroacetic acid modifier gradient). LCMS (ESI) m/z558.44 [M+H]⁺, t_(R)=1.88 min. (Gemini 5 u C18 110 Å, 50×4.60 mm 5micron column, 3.5 min, 2 ml/min, 5-100% MeCN/H₂O, 0.1% acetic acidmodifier gradient). R_(f)=0.25 (ethyl acetate) UV.

Example 22 Compound 22

Synthesis of Compound 22a:

A solution of 1d (865 mg, 2 mmol) in DCM (20 mL) was cooled in an icewater bath. Trimethylsilyl trifluoromethanesulfonate (667 mg, 3 mmol)was added dropwise at 0° C. under argon, and the resulting solution wasstirred at room temperature for 30 min. The reaction mixture wasevaporated to dryness and the resulting crude residue was dissolved indry acetonitrile (25 mL) under argon. Reaction mixture was stirred at 0°C., (S)-2-(tert-butoxycarbonyl(methyl)amino)-3-methylbutanoic aciddicyclohexylamine salt (908 mg, 2.2 mmol) and N,N-diisopropylethylamine(1.034 g, 8 mmol) were added followed by2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (1.065 g, 2.8 mmol). Reaction mixturewas stirred at room temperature for 48 h. The solvent was evaporated,the residue was dissolved in ethyl acetate (100 mL) and the solution waswashed twice with 20% water solution of citric acid (100 mL), water (100mL) and brine (100 mL), dried over magnesium sulfate, filtered andevaporated. The residue was purified by silica gel chromatography(gradient from 0-40% ethyl acetate+methanol (4/1) in hexane) to affordthe title compound as a white solid after evaporation (617 mg, 56%).

Synthesis of Compound 22b:

Under argon, a solution of diisopropylamine (2.51 g, 24.8 mmol) intetrahydrofuran (150 mL) was cooled in an ice water bath. A solution ofn-butyllithum in hexanes (2.5 M, 9.7 mL, 24 mmol) was added dropwiseover 2 min, and the resulting solution was stirred for 10 additionalmin. The solution was then cooled to −78° C. in a CO₂:acetone bath, andmethyl 2,2-dimethyl-3-oxobutanoate (3.2 g, 22 mmol) was added dropwiseover 30 sec. The solution was stirred for an additional 15 min, andN-phenyl-bis(trifluoromethanesulfonimide) (8.4 g, 23.5 mmol) was addedas a solution in tetrahydrofuran (20 mL) via cannula over 5 min, washingwith an additional portion of tetrahydrofuran (10 mL). The resultingsolution was stirred for 10 min and was removed from the cold bath.After stirring an additional 1 h, the reaction mixture was concentratedin vacuo and diluted with diethyl ether (150 mL). The organic phase waswashed with 1M aqueous sodium hydroxide (1×100 mL, 1×30 mL), dried overanhydrous magnesium sulfate, filtered, and concentrated to afford thetitle compound (6.2 g, 100%) as an amber liquid that was used withoutfurther purification.

Synthesis of Compound 22c:

A solution of 22b (6.2 g, 22 mmol) in anhydrous pyridine (11 mL, 140mmol) was heated to 70° C. After 18.5 h, the temperature was increasedto 90° C. After stirring for a total of 72 h, the reaction mixture waspartitioned between a stirred mixture of diethyl ether (100 mL) and 3 Maqueous HCl (100 mL). The phases were separated, and the organic layerwas washed with saturated aqueous sodium bicarbonate (75 mL), dried overmagnesium sulfate, filtered, and concentrated to afford the titlecompound (2.7 g, 97%) as a slightly brown liquid that was used withoutfurther purification.

Synthesis of Compound 22d:

Under argon, bis(cyclopentadienyl) zirconium chloride hydride (290 mg,1.1 mmol) was cooled in an ice water bath. A solution of 22c (1.4 g,11.1 mmol) and pinacolborane (2.4 mL, 16.5 mmol) in dichloromethane (3mL) was added by cannula, washing with an additional portion ofdichloromethane (2 mL). The resulting mixture was removed from the coldbath and was stirred for 72 h at room temperature. The reaction was thendiluted with ethyl acetate (50 mL), quenched with dropwise water (5 mL),and was further diluted with water (50 mL). The organic and aqueousphases were separated, and the aqueous phase was extracted with ethylacetate (30 mL). The combined organic phase was dried over anhydroussodium sulfate, filtered, and concentrated to afford a crude residuethat was purified by silica gel chromatography (5% to 15% ethyl acetatein hexanes) to afford the title compound (1.6 g, 57%) as a colorless oilthat crystallized on standing at −15° C.

Synthesis of Compound 22e:

Under argon, 20a (880 mg, 4.23 mmol), 22d (1.24 g, 4.88 mmol),PdCl₂(PCy₂(p-NMe₂Ph))₂ (173 mg, 0.21 mmol) and K₃PO₄ (2.64 g, 12.4 mmol)were dissolved in cyclopentyl methyl ether (11.9 mL) and water (5.1 mL).The resulting biphasic mixture was vigorously stirred in an oil bath at90° C. for 3.5 h, at which time the reaction was cooled to ambienttemperature and was diluted with ethyl acetate (50 mL) and water (40mL). The phases were separated, and the aqueous phase was extracted withethyl acetate (2×50 mL). The combined organic phase was dried overanhydrous magnesium sulfate, filtered, and concentrated to afford acrude residue that was purified by silica gel chromatography (25 to 60%ethyl acetate in hexanes) to afford the title compound as a yellow oil(1.07 g, 85%).

Synthesis of Compound 22f:

To a solution of 22e (600 mg, 2 mmol) in tetrahydrofuran (8 mL) wasadded methanol (4 mL), water (4 mL) and lithium hydroxide (96 mg, 4mmol). The resulting mixture was stirred at room temperature for 10 hand quenched with 1 M HCl (4.2 mL, 4.2 mmol). The resulting solution wasconcentrated to a crude residue which was co-distilled twice withtetrahydrofuran (20 mL), twice with dry acetonitrile (20 mL) and twicewith dry toluene (20 mL). The resulting white solid was dried under highvacuum overnight and it was used without further purification (735 mg,quantitative yield).

Synthesis of Compound 22g:

A solution of 17a (230 mg, 0.42 mmol) in dichloromethane (10 mL) wascooled in an ice water bath. Trimethylsilyl trifluoromethanesulfonate(128 mg, 0.575 mmol) was added dropwise at 0° C. under argon, and theresulting solution was stirred at room temperature for 30 min. Thereaction mixture was evaporated to dryness and the resulting cruderesidue was dissolved in dry acetonitrile (20 mL) under argon. Reactionmixture was stirred at 0° C., 22f (128 mg, 0.38 mmol) andN,N-diisopropylethylamine (198 mg, 1.53 mmol) were added followed by2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (218 mg, 0.58 mmol). Reaction mixturewas stirred at 50° C. for 5 days. The solvent was evaporated, theresidue was dissolved in ethyl acetate (50 mL) and the solution waswashed twice with 20% water solution of citric acid (50 mL), water (50mL) and brine (50 mL), dried over magnesium sulfate, filtered andevaporated. The residue was purified by silica gel chromatography(gradient from 0-40% ethyl acetate+methanol (4/1) in hexane) to affordafter evaporation the title compound as a white solid (88 mg, 32%).

Synthesis of the Title Compound:

To a solution of 22g (45 mg, 0.06 mmol) in tetrahydrofuran (2 mL) wasadded methanol (1 mL), water (1 mL) and lithium hydroxide (2.9 mg, 0.12mmol). The mixture was stirred for 2 h at ambient temperature and wasquenched with aqueous 1 M HCl (0.14 mL, 0.14 mmol). The resultingsolution was concentrated to a crude residue which was co-distilledtwice with tetrahydrofuran (5 mL), twice with dry acetonitrile (5 mL)and twice with dry toluene (5 mL). The resulting white solid was driedunder high vacuum overnight and it was used without further purification(40 mg, quantitative yield). Into oven dried, argon purged flask wereplaced 2-methyl-6-nitrobenzoic anhydride (83 mg, 0.24 mmol),4-dimethylformamide (88 mg, 0.72 mmol) and dry 1,2-dichloroethane (100mL). The resulting solution was heated at 70° C., and the crude productfrom the previous step was added dropwise via syringe as a solution indry N,N-dimethylformamide (5 mL) over 12 h. An additional portion of dryN,N-dimethylformamide (2×1 ml) was used to complete the quantitativetransfer. After stirring for additional 8 h at 70° C., the reactionmixture was transferred to separatory funnel and washed with water (100mL, 5 mL of brine was added to support the separation). The aqueousphase was extracted with dichloromethane (50 mL). Combined organicextracts were washed with brine (50 mL) and dried over magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingresidue was dissolved in ethyl acetate (100 mL) and was washed withwater (100 mL, 5 mL of brine was added to support the separation). Theaqueous phase was extracted with ethyl acetate (50 mL). Combined organicextracts were washed with water (100 mL, 5 mL of brine was added tosupport the separation). Resulting aqueous phase was extracted withethyl acetate (50 mL). Combined organic extracts were washed with brine(50 mL) and dried over magnesium sulfate, filtered and concentratedunder reduced pressure. The residue was purified by silica gelchromatography (gradient from 0-40% ethyl acetate+methanol (4/1) inhexane) to afford after evaporation the title compound (6 mg, 18%) as acolorless oil. ¹H NMR (400 MHz, CD₃OD): δ 9.05 (s, 1H), 7.95 (d, J=8.5Hz, 1H), 7.70 (s, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.20 (s, 1H), 6.76 (d,J=16.2 Hz, 1H), 6.28 (d, J=16.2 Hz, 1H), 5.93 (m, 1H), 5.27-5.22 (m,1H), 5.14 (m 1H), 4.82 (m 1H), 4.29 (m 1H), 3.70 (m, 1H), 3.38 (m, 1H),2.77 (s, 3H), 2.63 (m, 1H), 2.09 (m, 2H), 1.93 (m, 2H), 1.86 (m, 1H),1.79 (m, 2H), 1.59 (t, J=7.4 Hz, 6H), 1.45 (m, 2H), 0.85 (d, J=6.6 Hz,3H), 0.76 (d, J=6.4 Hz, 3H). LC/MS (m/z) 564 (100%) [M+H]⁺; R_(T)=3.14min.; purity >95% (Phenomenex Gemin 5 micron C18, 30×4.6 mm, 5 to 100%acetonitile in water over 6 min, 2 mL/min, 0.05% formic acid modifier).TLC Rf=0.43, 5% methanol in dichloromethane.

Example 23 Compound 23

Synthesis of Compound 23a:

Under argon, (S)-methyl 2-hydroxy-3-methylbutanoate (578 mg, 4.37 mmol)was dissolved in tetrahydrofuran (3.7 mL) and N,N-dimethylformamide (2.5mL). The solution was cooled in an ice water bath. Sodiumbis(trimethylsilyl)amide was added as a 1.0 M solution intetrahydrofuran (3.7 mL, 3.7 mmol) dropwise over 1.5 min. After anadditional 3.5 min, 5-bromopent-1-ene was added in one portion. Thereaction was stirred for 2 min and was removed from the cold bath. After2 h, the reaction was quenched with saturated aqueous ammonium chloride(10 mL) and was diluted with ethyl acetate (20 mL), water (10 mL), and0.1 N HCl (5 mL). The phases were separated, and the aqueous phase wasextracted with ethyl acetate (20 mL). The combined organic phase wasdried over sodium sulfate, filtered, and concentrated to afford a cruderesidue that was purified by silica gel chromatography (0 to 10% ethylacetate in hexanes) to afford the title compound (290 mg, 33%) as acolorless liquid

Synthesis of Compound 23b:

To a solution of 23a (155 mg, 0.774 mmol) in tetrahydrofuran (3 mL) wasadded water (1 mL) and lithium hydroxide monohydrate (165 mg, 3.93mmol). The reaction was stirred 18 h at which time methanol (1 mL) wasadded. The reaction was stirred for an additional 24 h and was thenconcentrated by 50% in vacuo and diluted with water (20 mL), ethylacetate (20 mL) and 3 N aqueous HCl (15 mL). The phases were separated,and the aqueous phase was extracted with ethyl acetate (2×20 mL). Thecombined organic phase was dried over anhydrous sodium sulfate filtered,and concentrated to afford (S)-3-methyl-2-pent-4-enyloxy-butyric acid(115 mg, 80%) which was used without further purification. Under argon,Id (240 mg, 0.55 mmol) was dissolved in dichloromethane (10 mL). Thesolution was cooled in an ice water bath and trimethylsilyltrifluoromethanesulfonate (209 mg, 0.94 mmol) was added. The reactionwas stirred for 1 h and was quenched by the addition ofN,N-diisopropylethylamine (296 mg, 2.3 mmol). The mixture wasconcentrated in vacuo and dissolved in methanol. After stirring for 10min, the solution was concentrated in vacuo and was twice dissolved inand concentrated from toluene (10 mL portions). The resulting cruderesidue was used without further purification.(S)-3-methyl-2-pent-4-enyloxy-butyric acid (0.62 mmol) and(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloroethyl ester (ca. 0.55 mmol) were dissolved inN,N-dimethylformamide (5 mL). N,N-diisopropylethylamine (192 mg, 1.5mmol) was added, and the solution was cooled in an ice water bath.(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate (320 mg, 0.75 mmol) was added in one portion, andthe reaction was stirred for 15 min. The reaction was then removed fromthe cold bath and let warm to ambient temperature. After stirringovernight, the reaction was diluted with ethyl acetate (30 mL) andsaturated aqueous sodium bicarbonate (30 mL). The phases were separated,and the organic phase was washed with a mixture of 0.2 N aqueous HCl (30mL) and brine (5 mL) followed by half-saturated brine (30 mL) and brine(30 mL). The organic phase was dried over anhydrous sodium sulfate,filtered, and concentrated to afford a crude residue that was purifiedby silica gel chromatography (30% to 60% ethyl acetate in hexanes) toafford the title compound (157 mg, 57%) as an amorphous residue.

Synthesis of Compound 23c:

To a solution of 23b (157 mg, 0.31 mmol) in tetrahydrofuran (5.3 mL) wasadded water (2.6 mL), ammonium acetate (365 mg, 4.7 mmol) and zinc dust(460 mg, 7.0 mmol). The resulting mixture was stirred vigorously at roomtemperature for 24 h, and the reaction was then heated to 50° C. Afteran additional 3.5 h, additional zinc powder (240 mg, 3.7 mmol) wasadded. The reaction was stirred at 50° C. for 2 h and was then cooled to35° C. and stirred for an additional 18.5 h. The mixture was filteredthrough Celite with ethyl acetate (50 mL) and was washed with a mixtureof 2 N aqueous HCl (30 mL) and brine (15 mL). The layers were separated,and the aqueous phase was extracted with ethyl acetate (2×30 mL). Thecombined organic phase was dried over sodium sulfate, filtered, andconcentrated. The resulting crude residue was twice taken up in andconcentrated from toluene (10 mL), and the crude(S)-1-[(S)-2-((S)-3-methyl-2-pent-4-enyloxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid was used without further purification.(S)-1-[(S)-2-((S)-3-Methyl-2-pent-4-enyloxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (74 mg, 0.37 mmol) and 4-dimethylaminopyridine (45.5 mg, 0.37 mmol)were dissolved in dichloromethane (5 mL).N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (78 mg,0.41 mmol) was added in one portion. The reaction was stirred 44 h atroom temperature and was diluted with ethyl acetate (50 mL), water (25mL), saturated aqueous sodium bicarbonate (25 mL), and brine (25 mL).The phases were separated, and the aqueous phase was extracted withethyl acetate (50 mL). The combined organic phase was dried overanhydrous sodium sulfate, filtered, and concentrated to afford a cruderesidue that was purified by silica gel chromatography (50 to 80 to 100%ethyl acetate in hexanes) to afford the title compound (118 mg, 69% over2 steps) as a white foam.

Synthesis of the Title Compound:

23c (53 mg, 0.096 mmol) was dissolved in toluene (30 mL) under argon.The solution was sparged with argon with stirring for 10 min.Grubbs-Hoveyda second generation metathesis catalyst (9.0 mg, 0.014) wasthen added as a solution in argon-sparged toluene (2.0 mL). The reactionwas heated to 100° C. in a preheated oil bath and was stirred for 35min. The reaction was concentrated in vacuo to ˜6.5 mL final volume andwas loaded directly onto a silica gel column. Elution with 60% to 85% to100% ethyl acetate in hexanes provided 30 mg of impure material that wasre-purified by reverse-phase HPLC (C18, 15% to 100% acetonitrile inwater, 0.1% trifluoroacetic acid) to provide the title compound (17.8mg, 35%) as its trifluoroacetic acid salt as an amorphous solid. ¹H NMR(400 MHz, CD₃OD) δ 9.56 (s, 1H), 8.53 (s, 1H), 8.39 (d, J=8.8 Hz, 1H),8.20 (s, 1H), 7.87 (d, J=8.6 Hz, 1H), 7.60-7.45 (m, 1H), 6.73 (d, J=16.1Hz, 1H), 6.21 (q, J=6.7 Hz, 1H), 5.93-5.79 (m, 1H), 4.49-4.36 (m, 1H),3.84-3.73 (m, 1H), 3.61 (d, J=6.7 Hz, 1H), 3.59-3.45 (m, 2H), 2.86-2.70(m, 2H), 2.44-2.31 (m, 1H), 2.18-1.89 (m, 4H), 1.88-1.61 (m, 7H), 1.59(d, J=7.1 Hz, 3H), 1.04-0.95 (m, 6H) ppm. LCMS (m/z) 523.3 [M+H], R_(T)2.72 min.

Example 24 Compound 24

The trifluoroacetic acid salt of Compound 23 (7.9 mg, 0.012 mmol) wasdissolved in ethyl acetate (0.80 mL). 10% palladium on carbon (5.6 mg)was added in one portion, and the reaction vessel was flushed withhydrogen. The reaction mixture was stirred for 2.5 h under 1 atmhydrogen gas and was then filtered through a pad of Celite with ethylacetate. The filtrate was concentrated and the resulting crude residuewas purified by reverse-phase HPLC (C18, 15% to 100% acetonitrile inwater, 0.1% trifluoroacetic acid) to provide the title compound (4.2 mg,60%) as its trifluoroacetic acid salt as an amorphous solid followinglyophilization. ¹H NMR (400 MHz, CD₃OD) δ 9.63 (s, 1H), 8.42 (d, J=8.8Hz, 1H), 8.26 (s, 1H), 7.96 (s, 1H), 7.91 (d, J=8.8 Hz, 1H), 6.16 (q,J=6.5 Hz, 1H), 5.79-5.68 (m, 1H), 3.91-3.83 (m, 1H), 3.46-3.40 (m, 1H),3.40 (d, J=5.7 Hz, 1H), 3.26-3.09 (m, 4H), 2.06-1.89 (m, 6H), 1.84 (m,1H), 1.73-1.59 (m, 7H), 1.52 (d, J=6.9 Hz, 3H), 1.52 (m, 1H), 0.93 (s,3H), 0.91 (s, 3H) ppm. LCMS (m/z) 525.4 [M+H], R_(T)=2.51 min.

Example 25 Compound 25

Compound 25a. 2,2-Dimethyl-but-3-en-1-ol

A solution of 2,2-dimethyl-but-3-enoic acid (570 mg, 5 mmol) andtriethylamine (505 mg, 0.7 mL, 5 mmol) in tetrahydrofuran (15 mL) wasstirred at −5° C. under nitrogen. A solution of methyl chloroformate(473 mg, 0.4 mL, 5 mmol) in tetrahydrofuran (5 mL) was added and thereaction mixture was stirred at −5° C. for 30 min. The reaction mixturewas filtered, the residue was washed with tetrahydrofuran and thefiltrate was evaporated. The residue was dissolved in tetrahydrofuran (5mL), water (5 mL) was added and the reaction mixture was stirred at 0°C. Sodium borohydride (190 mg, 5 mmol) was added portion-wise and thereaction mixture was stirred at 0° C. for 15 min and then at roomtemperature for 30 min. Saturated ammonium chloride solution (25 mL) wasadded and the mixture was extracted with diethyl ether. The organicextracts were combined, washed with brine, dried over anhydrousmagnesium sulfate, filtered and evaporated. The residue was purified bysilica gel chromatography using a gradient of iso-hexanes/diethyl ether10:1 to 5:1 to afford the title compound (569 mg, 95%) as an oil. ¹H NMR(300 MHz, CDCl₃) δ1.05 (s, 6H), 1.33 (t, J=6.5 Hz, 1H), 3.36 (d, J=6.5Hz, 2H), 5.09 (dd, J=17.4, 1.3 Hz, 1H), 5.12 (dd, J=10.9, 1.3 Hz, 1H),5.79 (dd, J=17.4, 10.9 Hz, 1H).

Compound 25b: 1-(7-Bromo-quinolin-2-yl)-ethanone

To a stirred slurry of 7-bromo-2-chloro-quinoline (obtained fromAstaTech, Inc.), (8.10 g, 33.4 mmol) and sodium iodide (50.0 g, 334mmol) in acetonitrile (27 mL) was slowly added acetyl chloride (3.56 mL,50.0 mmol). The flask was stoppered and sealed and heated at 80° C. for3 h before being allowed to cool. The mixture was treated sequentiallywith 10% w/w aqueous potassium carbonate solution (80 mL), 5% w/waqueous sodium sulfite solution (80 mL) and saturated aqueous sodiumthiosulfate solution (80 mL) and the mixture extracted withdichloromethane (2×). The combined organic extracts were dried oversodium sulfate, filtered and evaporated to give a crude7-bromo-2-iodo-quinoline. To the quinoline was addedtributyl(1-ethoxyvinyl)tin (13.6 mL, 40.1 mmol), 1,4-dioxane (67 mL) andbis(triphenylphosphine)palladium(II) dichloride (2.37 g, 3.34 mmol) andthe reaction mixture heated at 100° C. for 5 h before being allowed tocool. 2 M aqueous hydrochloric acid (67 mL) was added and the reactionstirred for 1 h. The mixture was filtered and the solids washed withethyl acetate and the filtrate evaporated to remove organics. Theresidue was extracted with ethyl acetate (3×) and the combined organicextracts were dried over sodium sulfate, filtered and evaporated. Theproduct was purified on silica gel doped with 10% w/w potassiumcarbonate eluting with a gradient of 0% to 6% ethyl acetate iniso-hexanes to afford the title compound (5.5 g, 66%) as a white solid.¹H NMR (300 MHz, CDCl₃) δ2.86 (s, 3H), 7.72-7.80 (m, 2H), 8.21 (ABq,Δδ_(AB)=0.11, J_(AB)=8.5 Hz, 2H), 8.42 (s, 1H). LCMS (m/z) 250.1/252.1[M+H], Tr=2.90 min.

Compound 25c: (R)-1-(7-Bromo-quinolin-2-yl)-ethanol

Dichloro (p-cymene) ruthenium(II) dimer (61 mg, 0.100 mmol) and(1R,2R)-(−)—N-p-tosyl-1,2-diphenylethylenediamine (88 mg, 0.012 mmol)was suspended in degassed water (40 mL) and the mixture was degassedwith nitrogen for 5 min. The mixture was stirred at 70° C. undernitrogen for 90 min. The resulting turbid orange mixture was allowed tocool to room temperature. 1-(7-Bromo-quinolin-2-yl)-ethanone (5.00 g, 20mmol) in degassed tetrahydrofuran (40 mL) was added followed by sodiumformate (6.8 g, 100 mmol) and the reaction mixture was degassed withnitrogen for 5 min. The reaction mixture was vigorously stirred at 40°C. for 4 h and allowed to cool. It was then diluted with ethyl acetateand water and the organic layer was separated, washed with water andbrine, dried over sodium sulfate, filtered and evaporated. The residuewas purified by silica gel chromatography using a gradient of 0% to 30%ethyl acetate in iso-hexanes to afford the title compound (4.96 g, 98%)as an off-white solid. ¹H NMR (300 MHz, CDCl₃) δ1.59 (d, J=6.7 Hz, 3H),4.85 (d, J=4.5 Hz, 1H), 5.04-5.07 (m, 1H), 7.40 (d, J=8.5 Hz, 1H), 7.64(dd, J=8.7, 1.6 Hz, 1H), 7.71 (d, J=8.7 Hz, 1H), 8.15 (d, J=8.5 Hz, 1H),8.28 (d, J=1.6 Hz, 1H). LCMS (m/z) 252.1/254.1 [M+H], Tr=1.74 min.

Compound 25d: Acetic acid (R)-1-(7-bromo-quinolin-2-yl)-ethyl ester

To a solution of 25c (1.00 g, 3.97 mmol) and triethylamine (1.65 mL,11.9 mmol) in anhydrous dichloromethane at 0° C., was added aceticanhydride (0.75 mL, 7.93 mmol) and 4-(dimethylamino)pyridine (24 mg,0.197 mmol). The reaction mixture was stirred and allowed to warm toroom temperature. After 1.5 h water (100 mL) was added and the layersseparated. The aqueous phase was re-extracted with dichloromethane(2×100 mL) and the combined organics were washed with brine (100 mL),dried over magnesium sulfate, filtered and concentrated in vacuo to givea crude residue. This was purified by silica gel chromatography usingiso-hexanes (66 mL) then iso-hexanes/ethyl acetate 95:5 (300 mL), theniso-hexanes/ethyl acetate 9:1 (1066 mL) to yield 25d (1.16 g, 99%) as acolorless oil. ¹H NMR (300 MHz, CD₃OD) δ1.65 (d, J=6.7 Hz, 3H), 2.16 (s,3H), 5.98 (q, J=6.7 Hz, 1H), 7.63 (d, J=8.5 Hz, 1H), 7.72 (dd, J=8.7,1.8 Hz, 1H), 7.87 (d, J=8.7 Hz, 1H), 8.23 (d, J=1.8 Hz, 1H), 8.37 (d,J=8.5 Hz, 1H). LCMS (m/z) 293.9/296.0 [M+H], Tr=2.58 min.

Compound 25e. Acetic acid(R)-1-[7-((E)-4-hydroxy-3,3-dimethyl-but-1-enyl)-quinolin-2-yl]-ethylester

A solution of 2,2-dimethyl-but-3-en-1-ol (40 mg, 0.4 mmol) and 25d (118mg, 0.4 mmol) in 1,4-dioxane (5 mL) was stirred at room temperatureunder nitrogen. Palladium(II) acetate (18 mg, 0.08 mmol) andtri(o-tolyl)phosphine (24 mg, 0.08 mmol) were added followed by asolution of N,N-dicyclohexylmethylamine (125 mg, 0.14 mL, 0.64 mmol) in1,4-dioxane (1 mL) and the reaction mixture was heated at 100° C. for 90min. Additional palladium(II) acetate (9 mg, 0.04 mmol),tri(o-tolyl)phosphine (12 mg, 0.04 mmol) were added and the reactionmixture was heated at 100° C. for 3 h. The reaction mixture was cooledto room temperature. Ethyl acetate was added and the mixture was washedwith saturated sodium hydrogen carbonate solution and brine. The organicsolution was filtered through a hydrophobic frit and the filtrate wasevaporated. The residue was purified by silica gel chromatography usingiso-hexanes/ethyl acetate 3:1 to 1:1 to afford 25e (85 mg, 68%) as agum. ¹H NMR (300 MHz, CDCl₃) δ1.20 (s, 6H), 1.43 (t, J=6.3 Hz, 1H), 1.69(d, J=6.7 Hz, 3H), 2.18 (s, 3H), 3.50 (d, J=6.3 Hz, 2H), 6.06 (q, J=6.7Hz, 1H), 6.52 (ABq, Δδ_(AB)=0.18, J_(AB)=16.0 Hz, 2H), 7.42 (d, J=8.5Hz, 1H), 7.62 (dd, J=8.5, 1.6 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 8.03 (brs, 1H), 8.11 (d, J=8.5 Hz, 1H). LCMS (m/z) 314.1 [M+H], Tr=2.21 min.

Compound 25f. Acetic acid(R)-1-[7-((E)-3,3-dimethyl-4-oxo-but-1-enyl)-quinolin-2-yl]-ethyl ester

A solution of 25e (78 mg, 0.25 mmol) in dichloromethane (5 mL) wasstirred 0° C. under nitrogen. Dess-Martin periodinane (106 mg, 0.25mmol) was added and the reaction mixture was stirred at 0° C. for 60min. Additional Dess-Martin periodinane (22 mg, 0.05 mmol) was added andthe reaction mixture was stirred at 0° C. for 30 min and then at roomtemperature for 15 min. The reaction mixture was diluted withdichloromethane and the mixture was washed with saturated sodiumhydrogen carbonate solution, water and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography usingiso-hexanes/ethyl acetate 4:1 to 1:1 to afford 25f (85 mg, 68%) as agum. ¹H NMR (300 MHz, CDCl₃) δ1.37 (s, 6H), 1.69 (d, J=6.7 Hz, 3H), 2.18(s, 3H), 6.06 (q, J=6.7 Hz, 1H), 6.40 (d, J=16.3 Hz, 1H), 6.65 (d,J=16.3 Hz, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.62 (dd, J=8.5, 1.2 Hz, 1H),7.76 (d, J=8.5 Hz, 1H), 8.04 (br s, 1H), 8.13 (d, J=8.5 Hz, 1H), 9.52(s, 1H). LCMS (m/z) 312.1 [M+H], Tr=2.65 min.

Compound 25g.(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of 20c (133 mg, 0.25 mmol) in dichloromethane (5 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (111 mg, 0.09 mL, 0.5 mmol) was added dropwiseand the reaction mixture was stirred at 0° C. for 1 h. Cold saturatedaqueous sodium hydrogen carbonate solution (10 mL) was added and themixture was stirred at 0° C. for 15 min. Dichloromethane was added andthe organic layer was separated, washed with brine, dried over anhydrousmagnesium sulfate and evaporated to afford(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (0.25 mmol) which was used withoutfurther purification. A solution of(S)-1-[(S)-2-((S)-2-amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (0.25 mmol) and 25f (66 mg, 0.21 mmol)in dichloromethane (5 mL) was stirred at room temperature for 10 min.Sodium triacetoxyborohydride (55 mg, 0.26 mmol) and acetic acid (1 drop)was added and the reaction mixture was stirred at room temperature for 4h. Additional sodium triacetoxyborohydride (13 mg, 0.25 mmol) was addedand the reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was diluted with ethyl acetate and the solution waswashed with saturated sodium hydrogen carbonate solution, water andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using iso-hexanes/ethyl acetate 1:1 to 0:1 to afford 25g(84 mg, 46%) as a gum. ¹H NMR (300 MHz, CDCl₃) δ0.86 (d, J=6.9 Hz, 3H),0.96 (d, J=6.9 Hz, 3H), 1.20 (s, 3H), 1.29 (d, J=7.0 Hz, 3H), 1.31 (s,3H), 1.69 (d, J=6.7 Hz, 3H), 1.70-2.15 (m, 6H), 2.18 (s, 3H), 2.52-2.63(m, 2H), 2.82-2.86 (m, 2H), 3.66-3.73 (m, 1H), 3.83 (d, J=11.4 Hz, 1H),4.45-4.50 (m, 1H), 4.74 (d, J=12.0 Hz, 1H), 4.95 (d, J=12.0 Hz, 1H),5.28-5.38 (m, 1H), 6.06 (q, J=6.7 Hz, 1H), 6.52 (ABq, Δδ_(AB)=0.14,J_(AB)=16.0 Hz, 2H), 7.42 (d, J=8.5 Hz, 1H), 7.66 (d, J=8.5 Hz, 1H),7.74 (d, J=8.5 Hz, 1H), 7.93 (d, J=8.5 Hz, 1H), 8.00 (s, 1H), 8.11 (d,J=8.5 Hz, 1H). LCMS (m/z) 726.1/728.1 [M+H], Tr=2.08 min.

Compound 25

A solution of 25g (80 mg, 0.11 mmol) in tetrahydrofuran (3 mL) wasstirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (18.5 mg, 0.44 mmol) in water (1 mL) was added and thereaction mixture was stirred at 0° C. for 4 h. Hydrochloric acid (1 M,0.45 mL) was added and the solvent was evaporated. The residue wasco-evaporated with tetrahydrofuran/toluene (1:1, 4×) and the residue wastriturated with diethyl ether (2×) and dried to afford(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.11 mmol) as a white solid which was used crude in the nextreaction. LCMS (m/z) 554.2 [M+H], Tr=1.03 min.

A mixture of crude(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.11 mol), N,N-diisopropylethylamine (84 mg, 0.65 mmol) and4-dimethylaminopyridine (5 mg, 0.04 mmol) in tetrahydrofuran (130 mL)was stirred at room temperature under nitrogen.2-(1H-7-Azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (133 mg, 0.35 mmol) was added and thereaction mixture was stirred at room temperature for 2.5 h. The solventwas evaporated and the residue was partitioned between ethyl acetate andsaturated sodium hydrogen carbonate solution. The organic solution wasseparated and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography using ethyl acetate. Theresidue was triturated with diethyl ether and the resulting solid wasdried to afford Compound 25 (21 mg, 30%) as a white solid. ¹H NMR (300MHz, CD₃OD) δ□0.80 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H), 1.11 (s,3H), 1.25 (s, 3H), 1.62 (d, J=7.1 Hz, 3H), 1.67-1.71 (m, 3H), 1.74 (d,J=6.7 Hz, 3H), 1.88-2.05 (m, 2H), 2.64 (d, J=8.5 Hz, 1H), 2.70-2.80 (m,3H), 3.80-3.85 (m, 1H), 4.35-4.40 (m, 1H), 5.89-5.99 (m, 2H), 6.42 (ABq,Δδ_(AB)=0.15, J_(AB)=16.5 Hz, 2H), 7.41 (d, J=8.5 Hz, 1H), 7.61 (dd,J=8.5, 1.5 Hz, 1H), 7.82 (d, J=8.5 Hz, 1H), 8.01 (br s, 1H), 8.22 (d,J=8.5 Hz, 1H). LCMS (m/z) 536.2 [M+H], Tr=1.47 min.

Example 26 Compound 26

Compound 26a.(S)-1-{(S)-2-[(S)-2-(tert-Butoxycarbonyl-methyl-amino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of 1d (1.08 g, 2.5 mmol) in dichloromethane (35 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (682 mg, 0.55 mL, 3.1 mmol) was added and thereaction mixture was stirred at 0° C. for 45 min.N,N-diisopropylethylamine (1.29 g, 1.73 mL, 10 mmol) was added and thesolvent was evaporated to afford crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (2.5 mmol), which was used in the next step.A solution of crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (2.5 mmol) in acetonitrile (30 mL) wasstirred at 0° C. under nitrogen.(S)-2-(tert-Butoxycarbonyl-methyl-amino)-3-methyl-butyric acid (635 mg,2.75 mmol, obtained from Sigma-Aldrich Inc.) andN,N-diisopropylethylamine (1.29 g, 1.7 mL, 10 mmol) was added followedby 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (1.33 g, 3.5 mmol) and the reactionmixture was stirred at room temperature for 18 h. The solvent wasevaporated and the residue was dissolved in ethyl acetate. The solutionwas washed with water, hydrochloric acid (2 M), water, saturated sodiumhydrogen carbonate solution, water and brine, dried over anhydroussodium sulfate, filtered and evaporated. The residue was purified bysilica gel chromatography using a gradient of iso-hexanes/ethyl acetate1:1 to 0:1 to afford the title compound (756 mg, 55%) as a white solid.¹H NMR (300 MHz, CDCl₃) δ0.88 (d, J=6.5 Hz, 3H), 0.97 (d, J=6.7 Hz, 3H),1.28 (d, J=6.7 Hz, 3H), 1.49-1.54 (m, 9H), 1.65-2.30 (m, 5H), 2.82 (s,3H), 2.82-2.94 (m, 1H), 3.62-4.18 (m, 3H), 4.30-4.45 (m, 1H), 4.73 (d,J=12.0 Hz, 1H), 4.95 (d, J=12.0 Hz, 1H), 5.29-5.39 (m, 1H), 6.62-6.77(m, 1H). LCMS (m/z) 545.2/547.1 [M+H], Tr=3.02 min.

Compound 26b.(S)-1-{(S)-2-[(S)-2-({(E)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enyl}-methyl-amino)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

Compound 26b was prepared in the same manner as compound 25g using 26ainstead of 20c in 42% yield. ¹H NMR (300 MHz, CDCl₃) δ0.90 (d, J=6.5 Hz,3H), 1.05 (d, J=6.5 Hz, 3H), 1.16 (s, 3H), 1.19 (s, 3H), 1.33 (d, J=6.7Hz, 3H), 1.69 (d, J=6.7 Hz, 3H), 1.70-2.15 (m, 5H), 2.17 (s, 3H), 2.40(s, 3H), 2.57-2.62 (m, 3H), 2.82-2.89 (m, 1H), 3.66-3.73 (m, 1H), 3.83(d, J=11.4 Hz, 1H), 4.43-4.48 (m, 1H), 4.72 (d, J=11.8 Hz, 1H), 4.94 (d,J=11.8 Hz, 1H), 5.27-5.37 (m, 1H), 6.06 (d, J=6.7 Hz, 1H), 6.50 (ABq,Δδ_(AB)=0.04, J_(AB)=16.5 Hz, 2H), 6.77 (d, J=8.0 Hz, 1H), 7.41 (d,J=8.3 Hz, 1H), 7.6 (d, J=8.5 Hz, 1H), 7.73 (d, J=8.5 Hz, 1H), 7.95 (brs, 1H), 8.11 (d, J=8.3 Hz, 1H). LCMS (m/z) 740.2/742.3 [M+H], Tr=2.44min.

Compound 26

Compound 26 was prepared in the same manner as compound 25 using 26binstead of 25 g in 11% yield as a white solid. ¹H NMR (300 MHz, CD₃OD)δ□0.86 (d, J=6.5 Hz, 3H), 0.96 (d, J=6.5 Hz, 3H), 1.14 (s, 3H), 1.23 (s,3H), 1.44 (d, J=7.1 Hz, 3H), 1.67-1.72 (m, 2H), 1.75 (d, J=6.9 Hz, 3H),1.81-2.25 (m, 3H), 2.51 (s, 3H), 2.79-2.91 (m, 4H), 3.69-3.73 (m, 1H),4.25-4.30 (m, 1H), 5.47 (q, J=6.7 Hz, 1H), 5.97 (q, J=6.7 Hz, 1H), 6.54(ABq, Δδ_(AB)=0.06, J_(AB)=16.0 Hz, 2H), 7.42 (d, J=8.3 Hz, 1H), 7.55(d, J=8.3 Hz, 1H), 7.81 (d, J=8.5 Hz, 1H), 7.95 (br s, 1H), 8.21 (d,J=8.5 Hz, 1H). LCMS (m/z) 550.3 [M+H], Tr=1.50 min.

Examples 27 and 28 Compound 27 and 28

Compound 27a. Trifluoro-methanesulfonic acid 2-methyl-but-3-enyl ester

A solution of 2-methyl-but-3-en-1-ol (840 mg, 10 mmol) and 2,6-lutidine(1.5 g, 1.6 mL, 14 mmol) in dichloromethane (40 mL) was stirred at −78°C. under nitrogen. Trifluoromethanesulfonic anhydride (3.38 g, 2.0 mL,12 mmol) was added dropwise and the reaction mixture was stirred at −78°C. for 1 h. Water was added and the reaction mixture was warmed to 0° C.Additional water was added and the organic layer was separated, washedwith cold 1 M hydrochloric acid, water and brine, dried over anhydrousmagnesium sulfate, filtered and evaporated to afford the crude product(14 mmol) as a pale yellow oil, which darkened on standing. The crudeproduct was used immediately in the next step. ¹H NMR (300 MHz, CDCl₃)δ1.16 (d, J=6.9 Hz, 3H), 2.66-2.76 (m, 1H), 4.33-4.55 (m, 2H), 5.20 (dd,J=10.5, 0.9 Hz, 1H), 5.21 (dd, J=16.3, 0.9 Hz, 1H), 5.66-5.81 (m, 1H).

Compound 27b. (S)-3-Methyl-2-(2-methyl-but-3-enyloxy)-butyric acidmethyl ester

A solution of (S)-2-hydroxy-3-methyl-butyric acid methyl ester (1.32 g,10 mmol) in tetrahydrofuran (30 mL) was stirred at −10° C. undernitrogen. Sodium bis(trimethylsilyl)amide (6 mL, 12 mmol, 2 M intetrahydrofuran) was added dropwise and the reaction mixture was stirredat −10° C. for 10 min. Trifluoro-methanesulfonic acid2-methyl-but-3-enyl ester (2.18 g, 10 mmol) was added in one portion andthe reaction mixture was stirred at room temperature for 2 h.

Saturated ammonium chloride solution was added and the mixture wasextracted with ethyl acetate. The organic extracts were combined, washedwith water and brine, dried over anhydrous magnesium sulfate, filteredand evaporated. The residue was purified by silica gel chromatographyusing a gradient of iso-hexanes/diethyl ether 20:1 to 10:1 to afford thetitle compound (429 mg, 21% over 2 steps) as an oil, as a 1:1 mixture oftwo diastereoisomers. ¹H NMR (300 MHz, CDCl₃) δ0.96-0.99 (m, 6H), 1.05(d, J=6.9 Hz, 1.5H), 1.08 (d, J=6.6 Hz, 1.5H), 2.01-2.12 (m, 1H),2.45-2.56 (m, 1H), 3.06-3.21 (m, 1H), 3.44-3.60 (m, 2H), 3.76 (s, 3H),5.00-5.11 (m, 2H), 5.74-5.88 (m, 1H).

Compound 27c. (S)-3-Methyl-2-(2-methyl-but-3-enyloxy)-butyric acid

A solution of 27b (429 mg, 2.1 mmol) in tetrahydrofuran (10 mL) wasstirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (265 mg, 6.3 mmol) in water (5 mL) was added followed bymethanol (2 mL) and the reaction mixture was stirred at 0° C. for 30minutes and then at room temperature for 4 h. Additional lithiumhydroxide monohydrate (90 mg, 2.1 mmol) and methanol (3 mL) were addedand the reaction mixture was stirred at room temperature for 3 days. Theorganic solvent was evaporated and the solution was acidified to pH 2with 2 M hydrochloric acid and the mixture was extracted with ethylacetate. The organic extracts were combined, washed with water andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated to afford the title compound (200 mg, 54%)as a colorless oil and as a 1:1 mixture of two diastereoisomers. ¹H NMR(300 MHz, CDCl₃) δ 0.97-1.11 (m, 9H), 2.14-2.23 (m, 1H), 2.48-2.59 (m,1H), 3.37-3.50 (m, 2H), 3.70-3.72 (m, 1H), 5.11-5.19 (m, 2H), 5.70-5.88(m, 1H), 8.50-9.50 (br s, 1H). LCMS (m/z) 185.3 [M−H], Tr=2.33 min.

Compound 27d.(S)-2-{(E)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyricacid

A solution of 27c (200 mg, 1.07 mmol) and 25d (314 mg, 1.07 mmol) inacetonitrile (10 mL) was stirred at room temperature under nitrogen.Palladium(II) acetate (48 mg, 0.2 mmol), tri(o-tolyl)phosphine (61 mg,0.2 mmol) was added followed by N,N-dicyclohexylmethylamine (418 mg,0.46 mL, 2.14 mmol) and the reaction mixture was heated at 120° C. in amicrowave reactor for 40 minutes. Additional palladium(II) acetate (48mg, 0.2 mmol), tri(-tolyl)phosphine (61 mg, 0.2 mmol) was added and thereaction mixture was heated at 130° C. in a microwave reactor for 30minutes. The solvent was evaporated and the residue was partitionedbetween ethyl acetate and 1 M hydrochloric acid. The organic extractswere combined and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated to afford thecrude product (1.07 mmol) as a 1:1 mixture of two diastereoisomers whichwas used in the next step without further purification. LCMS (m/z) 400.2[M+H], Tr=2.75 min.

Compound 27e.(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of 1d (433 mg, 1.0 mmol) in dichloromethane (30 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (444 mg, 0.35 mL, 2.0 mmol) was added and thereaction mixture was stirred at 0° C. for 45 minutes.N,N-diisopropylethylamine (516 mg, 0.7 mL, 4.0 mmol) was added and thesolvent was evaporated and the residue was dried to afford crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (1.0 mmol), which was used in the next step.A solution of crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (1.0 mmol) in acetonitrile (30 mL) wasstirred at 0° C. under nitrogen. 27d (1.0 mmol) andN,N-diisopropylethylamine (516 mg, 0.7 mL, 4.0 mmol) was added followedby 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (530 mg, 1.4 mmol) and the reactionmixture was stirred at 0° C. for 30 minutes and then at room temperaturefor 2 h. The solvent was evaporated and the residue was dissolved inethyl acetate. The solution was washed with cold 1 M hydrochloric acid,water, saturated sodium hydrogen carbonate solution, water and brine.The organic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:1 to 1:2to afford the title compound (420 mg, 58% over 2 steps) as a whitesolid, as a 1:1 mixture of two diastereoisomers. LCMS (m/z) 713.3/715.3[M+H], Tr=3.48 min.

Compound 27 and 28

A solution of 27e (143 mg, 0.2 mmol) in tetrahydrofuran (10 mL) wasstirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (42 mg, 1.0 mmol) in water (2 mL) was added and the reactionmixture was stirred at 0° C. for 3 h and then at 10° C. for 3 h. 1 MHydrochloric acid (1 mL, 1.0 mmol) was added and the solvent wasevaporated. The residue was co-evaporated with tetrahydrofuran/toluene(1:1, 3×) and the residue was triturated with diethyl ether (2×) anddried to afford(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.2 mmol) as a pale yellow solid, as a mixture of twodiastereoisomers which was used crude in the next reaction. LCMS (m/z)541.2 [M+H], Tr=1.77 min. A suspension of crude(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.2 mmol) in tetrahydrofuran (200 mL) was stirred at roomtemperature under nitrogen. N,N-Diisopropylethylamine (129 mg, 0.18 mL,1.0 mmol) and 4-dimethylaminopyridine (8 mg, 0.06 mmol) was added andthe reaction mixture was stirred at room temperature for 5 minutes.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (107 mg, 0.28 mmol) was added and thereaction mixture was stirred at room temperature for 2 h. Additional2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (30 mg, 0.08 mmol) was added and thereaction mixture was stirred at room temperature for 2 h. The solventwas evaporated and the residue was suspended in ethyl acetate. Cold 1 Mhydrochloric acid was added and the organic layer was separated, washedwith water, saturated sodium hydrogen carbonate solution, water andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:4 to 0:1to afford a 1:1 mixture of two diastereoisomers. The mixture waspurified by reverse phase preparative HPLC eluting with acetonitrile(containing 0.1% formic acid)/water (containing 0.1% formic acid) 45:55.Fractions containing the pure diastereoisomers were combined andevaporated. The residues were partitioned between saturated sodiumhydrogen carbonate solution and ethyl acetate. The organic layer wasseparated, washed with water and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated toafford two diastereoisomers.

Example 27 Compound 27 (First eluting) Diastereoisomer 1: (12 mg, 12%)as a white solid. ¹H NMR (300 MHz, d₆-DMSO) δ 0.70 (d, J=6.5 Hz, 3H),0.93 (d, J=6.5 Hz, 3H), 0.97 (d, J=6.9 Hz, 3H), 1.54 (d, J=6.9 Hz, 3H),1.55-1.62 (m, 2H), 1.63 (d, J=6.7 Hz, 3H), 1.73-1.95 (m, 3H), 2.72-2.78(m, 1H), 3.25-3.30 (m, 1H), 3.45-3.52 (m, 1H), 3.65-3.72 (m, 2H),4.18-4.22 (m, 1H), 4.87 (d, J=12.2 Hz, 1H), 5.65-5.73 (m, 1H), 5.96 (q,J=6.7 Hz, 1H), 6.34 (dd, J=16.5, 5.5 Hz, 1H), 6.45 (d, J=16.5 Hz, 1H),7.48 (d, J=8.3 Hz, 1H), 7.58 (d, J=8.7 Hz, 1H), 7.80 (s, 1H), 7.87 (d,J=8.5 Hz, 1H), 8.28 (d, J=8.5 Hz, 1H), 8.47 (d, J=8.3 Hz, 1H). LCMS(m/z) 523.3 [M+H], Tr=2.30 min.

Example 28 Compound 28 (Second Eluting) Diastereosiomer 2: (12 mg, 12%)as a white solid. ¹H NMR (300 MHz, d₆-DMSO) 60.73 (d, J=6.7 Hz, 3H),0.93 (d, J=6.5 Hz, 3H), 1.10 (d, J=6.9 Hz, 3H), 1.49 (d, J=7.1 Hz, 3H),1.55-1.62 (m, 2H), 1.65 (d, J=6.9 Hz, 3H), 1.75-1.95 (m, 3H), 2.56-2.62(m, 1H), 2.72-2.82 (m, 1H), 3.64-3.73 (m, 3H), 4.18-4.32 (m, 1H), 4.84(d, J=12.1 Hz, 1H), 5.65-5.73 (m, 1H), 5.98 (q, J=6.7 Hz, 1H), 6.34 (dd,J=16.5, 5.2 Hz, 1H), 6.48 (d, J=16.5 Hz, 1H), 7.48 (d, J=8.5 Hz, 1H),7.61 (d, J=8.5 Hz, 1H), 7.75 (s, 1H), 7.87 (d, J=8.5 Hz, 1H), 8.28 (d,J=8.3 Hz, 1H), 8.43 (d, J=8.0 Hz, 1H). LCMS (m/z) 523.3 [M+H], Tr=2.34min.

Example 29 Compound 29

Compound 29a.(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid

To(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester 1e (502 mg, 1.16 mmol) suspended intetrahydrofuran (15 mL) and water (3 mL) was added lithium hydroxidemonohydrate (69 mg, 1.65 mmol). The reaction mixture was stirred for 45minutes and then hydrochloric acid (2 M, 0.75 mL) was added. Thereaction mixture was stirred for 5 minutes and then evaporated. Thecrude residue was used directly in the next step without furtherpurification.

Compound 29b.(S)-1-[(S)-2-((S)-2-Amino-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (R)-1-(7-bromo-quinolin-2-yl)-ethyl amide

18c (533 mg, 1.50 mmol) was dissolved in hydrochloric acid in1,4-dioxane (4 M, 7.5 mL) and methanol (7.5 mL). The reaction mixturewas stirred for 2 h and then evaporated. The residue was passed throughan SCX cartridge eluting with methanol and then 1 M ammonia in methanol.The basic fraction was collected and evaporated. The residue wasdissolved in dichloromethane (30 mL) and 23 mL of this solution wasadded to freshly prepared 29a (350 mg, 1.16 mmol). The reaction vesselwas cooled to 0° C. and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (616 mg, 1.62 mmol) andN,N-diisopropylethylamine (450 mg, 606 μL, 3.48 mmol) were added. Thereaction mixture was allowed to warm to ambient temperature and stirredfor 15 h. The mixture was diluted with dichloromethane and washedsuccessively with saturated sodium bicarbonate solution, water andsaturated ammonium chloride solution, dried over anhydrous sodiumsulfate, filtered and evaporated. The residue was purified on silicaeluting with ethyl acetate/methanol 1:0 to 19:1 to afford the titlecompound (205 mg, 33%) as a white foam. ¹H NMR (300 MHz, CDCl₃) δ0.85(d, J=6.9 Hz, 3H), 1.01 (d, J=6.9 Hz, 3H), 1.41 (d, J=6.9 Hz, 3H), 1.58(d, J=6.7 Hz, 3H), 1.60-1.81 (m, 2H), 2.04-2.16 (m, 1H), 2.20-2.29 (m,1H), 3.69-3.79 (m, 1H), 3.26 (br s, 1H), 3.43-3.54 (m, 1H), 3.72(heptet, J=6.4 Hz, 1H), 3.86 (d, J=11.8 Hz, 1H), 3.98 (d, J=3.1 Hz, 1H),4.47-4.56 (m, 1H), 5.27 (app pentet, J=6.9 Hz, 1H), 5.46 (dq, J=8.0, 6.9Hz, 1H), 7.13 (d, J=8.2 Hz, 1H), 7.39 (d, J=8.5 Hz, 1H), 7.65 (dd,J=8.5, 1.8 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 8.10 (d, J=6.7 Hz, 1H), 8.14(d, J=8.5 Hz, 1H), 8.43 (s, 1H). LCMS (m/z) 536.0 [M+H], Tr=2.21 min.

Compound 29c.(R)-4-Isopropyl-3-((E)-2-methyl-but-2-enoyl)-oxazolidin-2-one

A cooled (−20° C.) solution of tiglic acid (2.005 g, 20.031 mmol) inanhydrous tetrahydrofuran (50 mL) was sequentially treated withtriethylamine (6.1 mL, 44.068 mmol) and dropwise pivaloyl chloride (2.7mL, 22.034 mmol). After stirring at −20° C. for 30 min, lithium chloride(1.019 g, 24.037 mmol) and (R)-(+)-4-isopropyl-2-oxazolidinone (2.587 g,20.031 mmol) were added. The reaction mixture was allowed to slowly warmto room temperature, stirred for 3 days and was then quenched withsaturated ammonium chloride. The aqueous layer was extracted with ethylacetate (2×). The organics were combined, dried over anhydrous sodiumsulfate, filtered and the volatiles were removed in vacuo. The residuewas purified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to4:1 to afford the title compound (3.307 g, 78%) as a colorless solid. ¹HNMR (300 MHz, CDCl₃) δ0.92 (d, J=6.9 Hz, 3H), 0.94 (d, J=7.1 Hz, 3H),1.83 (d, J=6.9 Hz, 3H), 1.93 (s, 3H), 2.38 (d of heptet, J=6.9, 4.2 Hz,1H), 4.19 (dd, J=8.9, 4.6 Hz, 1H), 4.33 (app t, J=8.9 Hz, 1H), 4.49-4.58(m, 1H), 6.23 (q, J=7.1 Hz, 1H).

Compound 29d.(R)-4-Isopropyl-3-[(R)-2-methyl-2-(2-trimethylsilanyl-ethoxymethyl)-but-3-enoyl]-oxazolidin-2-one

Residual moisture in 29c (458.9 mg, 2.172 mmol) was removed byazeotroping with anhydrous toluene. A cooled (−78° C.) solution of dried29c in anhydrous toluene (10 mL) was treated dropwise with a solution ofsodium bis(trimethylsily)amide in toluene (0.6 M, 5.4 mL, 3.258 mmol).After stirring the yellow solution at −78° C. for 35 min,2-(trimethylsilyl)ethoxymethyl chloride (1.1 mL, 6.516 mmol) was added.After stirring at 0° C. for 3.5 h, the reaction was quenched withsaturated ammonium chloride. The aqueous layer was extracted withdichloromethane (2×). The organics were combined, filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to9:1 to afford the title compound (543.3 mg, 73%) as a colorless oil andas a 5:1 mixture of diastereoisomers. ¹H NMR (300 MHz, CDCl₃) δ0.01 (s,9H), 0.77-0.99 (m, 8H), 1.49 (s, 3H), 2.36 (d of heptet, J=6.9, 3.8 Hz,1H), 3.42-3.60 (m, 3H), 4.15-4.31 (m, 3H), 4.49-4.57 (m, 1H), 5.00 (d,J=17.6 Hz, 1H), 5.10 (d, J=10.7, 1H), 6.19 (dd, J=17.8, 10.7 Hz, 1H).LCMS (m/z) 364.1 [M+Na], Tr=3.32 min.

Compound 29e.(R)-3-((R)-2-Hydroxymethyl-2-methyl-but-3-enoyl)-4-isopropyl-oxazolidin-2-one

To a stirred solution of 29d (2.93 g, 8.58 mmol) in a mixture ofacetonitrile (86 mL) and water (1.72 mL) was added lithiumtetrafluoroborate (1 M in acetonitrile, 42.9 mL, 42.9 mmol). The mixturewas heated at reflux under nitrogen for 5 h and allowed to cool toambient temperature. More lithium tetrafluoroborate (1 M inacetonitrile, 8.6 mL, 8.6 mmol) was added and the mixture heated atreflux for a further 75 minutes and allowed to cool to ambienttemperature. The mixture was diluted with water/diethyl ether (2:3, 100mL) and the organic layer was separated and washed with water. Thecombined aqueous washes were back-extracted with diethyl ether and thecombined organic extracts dried over anhydrous sodium sulfate, filteredand evaporated to give the title compound (1.89 g, 91%) as a yellow oil.¹H NMR (300 MHz, CDCl₃) δ0.89 (d, J=6.9 Hz, 3H), 0.95 (d, J=6.9 Hz, 3H),1.35 (s, 3H), 2.36-2.47 (m, 1H), 2.56-2.70 (br s, 1H), 3.47-3.54 (m,1H), 3.91 (d, J=11.6 Hz, 1H), 4.22-4.31 (m, 2H), 4.47-4.53 (m, 1H), 4.97(d, J=17.8 Hz, 1H), 5.17 (d, J=10.7 Hz, 1H), 6.09 (dd, J=17.8, 10.7 Hz,1H). LCMS (m/z) 242.2 [M+H], Tr=1.82 min.

Compound 29f.(R)-3-[(R)-2-(tert-Butyl-dimethyl-silanyloxymethyl)-2-methyl-but-3-enoyl]-4-isopropyl-oxazolidin-2-one

To a stirred solution of 29e (660 mg, 2.74 mmol) andtert-butyldimethylsilyl chloride (533 mg, 3.56 mmol) inN,N-dimethylformamide (10 mL) at 0° C. under nitrogen was added in oneportion imidazole (821 mg, 12.1 mmol). The reaction was stirred at 0° C.for 15 minutes and then allowed to warm to ambient temperature andstirred for 5 h. The mixture was concentrated and then partitionedbetween saturated ammonium chloride solution and ether. The organiclayer was separated and washed with water (3×), dried over anhydroussodium sulfate, filtered and evaporated. The residue was purified onsilica gel chromatography eluting with iso-hexanes/ethyl acetate 1:0 to19:1 to afford the title compound (316 mg, 32%) as a colorless oil. ¹HNMR (300 MHz, CDCl₃) δ0.05 (s, 3H), 0.06 (s, 3H), 0.86-0.94 (m, 6H),0.89 (s, 9H), 1.45 (s, 3H), 2.27-2.40 (m, 1H), 3.71 (d, J=9.6 Hz, 1H),4.10-4.30 (m, 2H), 4.43 (d, J=9.6 Hz, 1H), 4.49-4.55 (m, 1H), 4.99 (d,J=17.8 Hz, 1H), 5.10 (d, J=9.6 Hz, 1H), 6.17 (dd, J=17.8, 10.9 Hz, 1H).LCMS (m/z) 356.2 [M+H], Tr=4.02 min

Compound 29g.(R)-2-(tert-Butyl-dimethyl-silanyloxymethyl)-2-methyl-but-3-enoic acid

To a stirred mixture of 29f (310 mg, 0.872 mmol) in tetrahydrofuran (4.4mL) and water (2.2 mL) at 0° C. was added hydrogen peroxide solution(30%, 494 mg, 449 μL, 4.36 mmol) followed by lithium hydroxidemonohydrate (73 mg, 1.74 mmol). The reaction mixture was stirred at 0°C. for 5 minutes then allowed to warm to ambient temperature and stirredfor 16 h. More tetrahydrofuran (1 mL) was added and the mixture cooledto 0° C. and lithium hydroxide monohydrate (37 mg, 0.87 mmol) was added.The reaction mixture was stirred at 0° C. for 40 minutes then allowed towarm to ambient temperature and stirred for 7 h. The reaction was thenquenched with sodium metabisulfite and stirred for 30 minutes. Themixture was acidified to pH<3 with hydrochloric acid (2 M) and extractedwith dichloromethane (3×). The combined organic extracts were dried overanhydrous magnesium sulfate, filtered and evaporated. The residue waspurified on silica gel chromatography eluting with iso-hexanes/ethylacetate 9:1 to afford the title compound (144 mg, 68%) as a colorlessoil. ¹H NMR (300 MHz, CDCl₃) δ0.12 (s, 6H), 0.93 (s, 9H), 1.31 (s, 3H),3.65 (d, J=9.6 Hz, 1H), 3.78 (d, J=9.6 Hz, 1H), 5.24 (d, J=17.6 Hz, 1H),5.24 (d, J=10.9 Hz, 1H), 5.96 (dd, J=17.4, 10.9 Hz, 1H).

Compound 29h.(S)-1-{(S)-2-[(S)-3-Methyl-2-((R)-2-methyl-2-tert-butyl-dimethyl-silanyloxymethyl-but-3-enoylamino)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (R)-1-(7-bromo-quinolin-2-yl)-ethyl amide

To a stirred solution 29b (504 mg, 0.944 mmol), 2-methyl-6-nitrobenzoicanhydride (649 mg, 1.89 mmol), 4-(dimethylamino)pyridine (46 mg, 0.378mmol), triethylamine (238 mg, 328 μL, 2.36 mmol) and powdered 4 Åmolecular sieves (˜1 g) in dichloromethane (10 mL) was added a solutionof 29g (346 mg, 1.42 mmol) in dichloromethane (4 mL). The reaction wasstirred under nitrogen for 91 h and then more 2-methyl-6-nitrobenzoicanhydride (325 mg, 0.90 mmol), 4-(dimethylamino)pyridine (23 mg, 0.189mmol) and a solution of 29g (173 mg, 0.71 mmol) in dichloromethane (2mL) was added and the reaction mixture stirred for a further 23 h. Thereaction mixture was filtered through Celite and cooled to 0° C. andice-cold water was added. The organic layer was separated and washedsequentially with ice-cold saturated ammonium chloride solution,ice-cold saturated sodium bicarbonate solution and saturated brine,dried over anhydrous sodium sulfate, filtered and evaporated andpurified on silica gel chromatography eluting with iso-hexanes/ethylacetate 1:1 to 0:1 to afford the title compound (244 mg, 34%) as a beigeoil. ¹H NMR (300 MHz, CDCl₃) δ0.05 (s, 3H), 0.06 (s, 3H), 0.88 (s, 9H),0.93 (d, J=6.9 Hz, 3H), 0.94 (d, J=6.9 Hz, 3H), 1.37 (d, J=6.5 Hz, 3H),1.42 (s, 3H), 1.61 (d, J=6.7 Hz, 3H), 1.54-1.71 (m, 2H), 1.91-2.01 (m,1H), 2.23-2.35 (m, 2H), 2.57-2.68 (m, 1H), 3.39-3.50 (m, 1H), 3.65 (d,J=9.5 Hz, 1H), 3.73 (d, J=12.1 Hz, 1H), 3.89 (d, J=9.6 Hz, 1H),4.51-4.60 (m, 1H), 5.02 (d, J=3.6 Hz, 1H), 5.21-5.43 (m, 4H), 6.04 (dd,J=17.4, 10.9 Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H),7.61-7.71 (m, 2H), 8.02 (d, J=6.5 Hz, 1H), 8.12 (d, J=8.5 Hz, 1H), 8.36(s, 1H). LCMS (m/z) 760.3, 762.3 [M+H], Tr=4.16 min.

Compound 29i

To solid palladium(II) acetate (14 mg, 0.060 mmol) andtri(o-tolyl)phosphine (20 mg, 0.060 mmol) was added a solution of 29h(49 mg, 0.060 mmol) in anhydrous 1,4-dioxane (18.4 mL), followed byN,N-dicyclohexylmethylamine (25 mg, 27 μL, 0.128 mmol) and anhydrousacetonitrile (0.2 mL). The mixture was heated at 120° C. in a microwavereactor for 25 minutes. Five batches of equivalent amount were reactedand combined and diluted with ethyl acetate, washed with saturatedammonium chloride solution (2×), brine, dried over anhydrous sodiumsulfate, filtered and evaporated. The residue was purified on silica gelchromatography eluting with iso-hexanes/ethyl acetate 1:1 to 1:3 toafford the title compound (81 mg, 37%) as a yellow gum. LCMS (m/z) 680.3[M+H], Tr=3.75 min

Compound 29

Compound 29i (40 mg, 0.059 mmol) was dissolved in acetonitrile (20 mL)and hydrofluoric acid (48% in water, 117 mg, 102 μL, 5.88 mmol) wasadded. The reaction mixture was stirred for 4 h and more hydrofluoricacid (48% in water, 117 mg, 102 μL, 5.88 mmol) was added and stirringcontinued for a further 3.5 h. Solid sodium bicarbonate was added andthe mixture stirred for 5 minutes, filtered through a hydrophobic fritand the filtrate evaporated. The residue was purified by reverse phasepreparative HPLC using a gradient of acetonitrile/water 1:4 to 7:3 toafford the title compound (10.0 mg, 30%) as a white solid. ¹H NMR (300MHz, CD₃OD) δ1.01 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.7 Hz, 3H), 1.48-1.78(m, 2H), 1.56 (s, 3H), 1.60 (d, J=6.9 Hz, 3H), 1.66 (d, J=7.4H, 3H),1.91-2.00 (m, 1H), 2.10-2.19 (m, 2H), 2.63-2.75 (m, 1H), 3.62 (dd,J=11.4, 2.7 Hz, 1H), 3.65 (d, J=10.9 Hz, 1H), 3.95 (d, J=10.9 Hz, 1H),4.38-4.47 (m, 1H), 5.08 (q, J=6.6 Hz, 1H), 5.23 (d, J=9.2 Hz, 1H), 5.80(q, J=7.2 Hz, 1H), 6.24 (d, J=16.5 Hz, 1H), 6.57 (d, J=16.5 Hz, 1H),7.45 (d, J=8.5 Hz, 1H), 7.66 (dd, J=8.5, 1.3 Hz, 1H), 7.83 (s, 1H), 7.85(d, J=8.5 Hz, 1H), 8.25 (d, J=8.5 Hz, 1H). LCMS (m/z) 566.3 [M+H],Tr=1.79 min.

Example 30 Compound 30

To a stirred solution of compound 29 (40 mg, 0.059 mmol) in ethylacetate (6 mL) was added 10% palladium on carbon (12.5 mg, 0.012 mmol).The mixture was reacted under an atmosphere of hydrogen for 1 h and thenthe catalyst was filtered off. Fresh 10% palladium on carbon (12.5 mg,0.012 mmol) was added to the filtrate and the stirred mixture reactedunder an atmosphere of hydrogen for 1 h. The mixture was filteredthrough Celite and then evaporated. The residue was dissolved in ethylacetate (6 mL) and fresh 10% palladium on carbon (12.5 mg, 0.012 mmol)was added and the stirred mixture reacted under an atmosphere ofhydrogen for 2 h. The mixture was filtered through Celite andevaporated. The residue was dissolved in acetonitrile (20 mL) andhydrogen fluoride (48% in water, 117 mg, 102 μL, 2.82 mmol) was added.The reaction mixture was stirred for 2 h and more hydrogen fluoride (48%in water, 117 mg, 102 μL, 2.82 mmol) was added and stirred for a further2 h. Solid sodium hydrogen carbonate was added and the mixture stirredfor 5 min and then filtered through a hydrophobic frit. The mixture wasevaporated and dissolved in ethyl acetate (6 mL) and 10% palladium oncarbon (12.5 mg, 0.012 mmol) was added. The stirred mixture was reactedunder a hydrogen atmosphere for 7 h and then filtered through Celite andevaporated. The residue was purified by two consecutive reverse phasepreparative HPLCs using a gradient of acetonitrile/water 1:4 to 3:7 andthen acetonitrile/water 1:4 to 1:1 modified with 0.1% formic acid toafford the title compound (3.1 mg, 9%) as a white solid. ¹H NMR (300MHz, CD₃OD) δ1.04 (d, J=6.7 Hz, 3H), 1.09 (d, J=6.9 Hz, 3H), 1.41 (s,3H), 1.48-1.75 (m, 2H), 1.58 (d, J=6.7 Hz, 3H), 1.60 (d, J=7.1H, 3H),1.80-1.99 (m, 3H), 2.12-2.25 (m, 1H), 2.26-2.36 (m, 1H), 2.61-2.73 (m,1H), 2.74-2.90 (m, 1H), 2.96-3.11 (m, 1H), 3.52 (d, J=10.9 Hz, 1H), 3.60(dd, J=11.2, 2.3 Hz, 1H), 3.80 (d, J=10.9 Hz, 1H), 4.24-4.42 (m, 1H),4.81-4.94 (m, 1H), 5.10 (q, J=6.7 Hz, 1H), 5.94 (d, J=7.2 Hz, 1H), 7.42(d, J=8.5 Hz, 1H), 7.43 (dd, J=8.3, 1.6 Hz, 1H), 7.80 (d, J=8.5 Hz, 1H),7.85 (s, 1H), 8.24 (d, J=8.5 Hz, 1H). LCMS (m/z) 568.2 [M+H], Tr=1.94min.

Example 31 Compound 31

Compound 31a: (S)-3-Methyl-2-[2,2,2-trifluoro-ethylideneamino]-butyricacid methyl ester

A solution of (S)-2-amino-3-methyl-butyric acid methyl esterhydrochloride (6.72 g, 40 mmol) in water (50 mL) was partitioned betweensaturated sodium hydrogen carbonate solution and dichloromethane. Theorganic layer was separated, washed with water and brine, dried overanhydrous magnesium sulfate, filtered and evaporated to afford(S)-2-amino-3-methyl-butyric acid methyl ester (40 mmol). A solution of(S)-2-amino-3-methyl-butyric acid methyl ester (40 mmol),2,2,2-trifluoro-1-methoxy-ethanol (5.2 g, 3.8 mL, 40 mmol) and4-toluenesulfonic acid hydrate (100 mg) in toluene (125 mL) was heatedat reflux using a Dean-Stark apparatus for 3 h. The reaction mixture wascooled to room temperature and the solvent was evaporated. The residuewas purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 10:1 to 5:1 to afford the title compound (4.79g, 56%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ0.94 (d, J=7.1 Hz,3H), 0.97 (d, J=7.1 Hz, 3H), 2.32-2.44 (m, 1H), 3.73-3.77 (m, 4H), 7.67(q, J=3.3 Hz, 1H).

Compound 31b:(S)-2-((S)-2,2-Dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid methyl ester

A solution of 31a (4.2 g, 20 mmol) and 3,3-dimethylallyl bromide (2.98g, 2.3 mL, 20 mmol) in tetrahydrofuran (30 mL) was stirred at roomtemperature under nitrogen. Zinc granules (2.6 g, 40 mmol) andchlorotrimethylsilane (10 drops) were added and the reaction mixture wasstirred at room temperature for 10 minutes and then heated at reflux for2 h. The reaction mixture was cooled to room temperature. Saturatedammonium chloride solution was added and the mixture was extracted withdiethyl ether. The organic layer was separated, washed with brine, driedover anhydrous magnesium sulfate, filtered and evaporated. The residuewas purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 50:1 to 40:1 to afford the title compound(1.41 g, 25%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ0.91 (d,J=6.7 Hz, 3H), 0.97 (d, J=6.7 Hz, 3H), 1.16 (s, 6H), 1.52-1.55 (m, 1H),1.81-1.92 (m, 1H), 2.76-2.85 (m, 1H), 3.20 (dd, J=9.8, 7.1 Hz, 1H), 3.72(s, 3H), 5.09 (d, J=17.4 Hz, 1H), 5.11 (d, J=10.8 Hz, 1H), 5.90 (dd,J=17.4, 10.8 Hz, 1H). LCMS (m/z) 282.1 [M+H], Tr=3.51 min.

Compound 31c:(S)-2-((S)-2,2-Dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid

A solution of 31b (1.12 g, 4.0 mmol) in tetrahydrofuran (40 mL) wasstirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (504 mg, 12.0 mmol) in water (8 mL) was added and thereaction mixture was stirred at 0° C. for 1 h and then at roomtemperature for 1 h. Methanol (5 mL) was added and the reaction mixturewas stirred at room temperature for 22 h. Additional lithium hydroxidemonohydrate (336 mg, 8.0 mmol) in water (2 mL) was added and thereaction mixture was stirred at room temperature for 24 h. The organicsolvent was evaporated and the solution was acidified to pH 2 with 2 Mhydrochloric acid and the mixture was extracted with ethyl acetate. Theorganic extracts were combined, washed with water and brine. The organicsolution was filtered through a hydrophobic frit and the filtrate wasevaporated to afford the title compound (770 mg, 72%) as a colorlessoil. ¹H NMR (300 MHz, CDCl₃) δ1.00 (d, J=6.7 Hz, 3H), 1.01 (d, J=6.9 Hz,3H), 1.20 (s, 6H), 1.95-2.04 (m, 1H), 2.87 (q, J=7.8 Hz, 1H), 3.37 (d,J=5.8 Hz, 1H), 5.12 (d, J=17.2 Hz, 1H), 5.16 (d, J=10.7 Hz, 1H), 5.90(dd, J=17.2, 10.7 Hz, 1H). LCMS (m/z) 268.2 [M+H], Tr=2.88 min.

Compound 31d:(S)-2-{(E)-(S)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyricacid

A solution of 31c (267 mg, 1.0 mmol) and 25d (294 mg, 1.0 mmol) inacetonitrile (10 mL) was stirred at room temperature under nitrogen.Palladium(II) acetate (45 mg, 0.2 mmol), tri(o-tolyl)phosphine (61 mg,0.2 mmol) were added followed by N,N-dicyclohexylmethylamine (390 mg,0.43 mL, 2.0 mmol) and the reaction mixture was heated at 120° C. in amicrowave reactor for 30 minutes. The solvent was evaporated and theresidue was partitioned between ethyl acetate and 1 M hydrochloric acid.The organic extracts were combined and washed with brine. The organicsolution was filtered through a hydrophobic frit and the filtrate wasevaporated to afford the crude product (640 mg, 1.0 mmol) as a yellowwaxy solid which was used in the next step without further purification.LCMS (m/z) 481.1 [M+H], Tr=3.26 min.

Compound 31e:(S)-1-[(S)-2-((S)-2-{(E)-(S)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of 1d (433 mg, 1.0 mmol) in dichloromethane (30 mL) wasstirred at 0° C. under nitrogen. Trimethylsilyltrifluoromethanesulfonate (444 mg, 0.35 mL, 2.0 mmol) was added and thereaction mixture was stirred at 0° C. for 45 minutes.N,N-diisopropylethylamine (516 mg, 0.7 mL, 4.0 mmol) was added, thesolvent was evaporated and the residue was dried affording crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (1.0 mmol), which was used in the next step.A mixture of crude(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid2,2,2-trichloro-ethyl ester (1.0 mmol), N,N-diisopropylethylamine (516mg, 0.7 mL, 4.0 mmol) and crude 31d (1.0 mmol) in acetonitrile (30 mL)was stirred at 0° C. under nitrogen.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (530 mg, 1.4 mmol) was added and thereaction mixture was stirred at 0° C. for 30 minutes and then at roomtemperature for 2 h. The solvent was evaporated and the residue wasdissolved in ethyl acetate and the solution was washed with cold 1 Mhydrochloric acid, water, saturated sodium hydrogen carbonate solution,water and brine. The organic solution was filtered through a hydrophobicfrit and the filtrate was evaporated. The residue was purified by silicagel chromatography using a gradient of iso-hexanes/ethyl acetate 3:1 to0:1 to afford the title compound (292 mg, 37% over 2 steps) as a whitefoam. ¹H NMR (300 MHz, CDCl₃) δ0.95 (d, J=7.1 Hz, 3H), 0.97 (d, J=7.1Hz, 3H), 1.30 (d, J=6.7 Hz, 3H), 1.38 (s, 6H), 1.62-1.68 (m, 3H), 1.69(d, J=6.7 Hz, 3H), 1.90-1.96 (m, 2H), 2.18 (s, 3H), 2.18-2.21 (m, 1H),2.84-3.23 (m, 3H), 3.68-3.72 (m, 1H), 3.79 (d, J=11.1 Hz, 1H), 4.37-4.42(m, 1H), 4.72 (d, J=11.8 Hz, 1H), 4.96 (d, J=11.8 Hz, 1H), 5.31-5.40 (m,1H), 6.06 (q, J=6.7 Hz, 1H), 6.59 (ABq, Δδ_(AB)=0.07, J_(AB)=16.2 Hz,2H), 7.05 (d, J=8.0 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H), 7.66 (d, J=8.5 Hz,1H), 7.76 (d, J=8.5 Hz, 1H), 8.01 (s, 1H), 8.12 (d, J=8.5 Hz, 1H). LCMS(m/z) 794.2/796.2 [M+H], Tr=3.48 min.

Compound 31

A solution of 31e (200 mg, 0.25 mmol) in tetrahydrofuran (10 mL) wasstirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (53 mg, 1.25 mmol) in water (2 mL) was added and thereaction mixture was stirred at 0° C. for 5 h. 1 M Hydrochloric acid(1.25 mL, 1.25 mmol) was added and the solvent was evaporated. Theresidue was co-evaporated with tetrahydrofuran/toluene (1:1, 3×) and theresidue was triturated with diethyl ether (2×) and the resulting solidwas dried to afford(S)-1-[(S)-2-((S)-2-{(E)-(S)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.25 mmol) as a white solid which was used crude in the nextreaction. LCMS (m/z) 622.3 [M+H], Tr=1.97 min. A suspension of crude(S)-1-[(S)-2-((S)-2-{(E)-(S)-4-[2-((R)-1-hydroxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.25 mmol) in tetrahydrofuran (250 mL) was stirred at 0° C. undernitrogen. N,N-Diisopropylethylamine (161 mg, 0.22 mL, 1.25 mmol) and4-dimethylaminopyridine (20 mg) were added and the reaction mixture wasstirred at 0° C. for 5 minutes.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (133 mg, 0.35 mmol) was added and thereaction mixture was stirred at 0° C. for 30 minutes and then at roomtemperature for 3 h. Additional2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (95 mg, 0.25 mmol) was added and thereaction mixture was stirred at room temperature for 18 h. The solventwas evaporated and the residue was suspended in ethyl acetate. Cold 1 Mhydrochloric acid was added and the organic layer was separated, washedwith water, saturated sodium hydrogen carbonate solution, water andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 4:1 to 2:1to afford the title compound (50 mg, 33% over 2 steps) as a white solid.¹H NMR (300 MHz, CD₃OD) δ 0.74 (d, J=6.7 Hz, 3H), 1.00 (d, J=6.7 Hz,3H), 1.29 (s, 3H), 1.35 (s, 3H), 1.55-1.70 (m, 3H), 1.71 (d, J=6.7 Hz,3H), 1.72 (d, J=6.5 Hz, 3H), 1.87-1.97 (m, 2H), 2.66-2.74 (m, 1H), 2.90(d, J=9.4 Hz, 1H), 3.13 (q, J=7.8 Hz, 1H), 3.70-3.79 (m, 1H), 4.28-4.37(m, 2H), 5.84 (q, J=7.1 Hz, 1H), 5.91 (q, J=6.7 Hz, 1H), 6.33 (d, J=16.2Hz, 1H), 6.63 (d, J=16.2 Hz, 1H), 7.42 (d, J=8.7 Hz, 1H), 7.49 (dd,J=8.3, 1.6 Hz, 1H), 7.83 (d, J=8.5 Hz, 1H), 8.20-8.23 (m, 2H). LCMS(m/z) 604.3 [M+H], Tr=3.25 min.

Example 32 Compound 32

Compound 32a: (R)-3-Methyl-2-[2,2,2-trifluoro-ethylideneamino]-butyricacid methyl ester

Compound 32a was prepared in the same manner as compound 31a using(R)-2-amino-3-methyl-butyric acid methyl ester hydrochloride instead of(S)-2-amino-3-methyl-butyric acid methyl ester hydrochloride in 64%yield. ¹H NMR (300 MHz, CDCl₃) δ0.94 (d, J=7.1 Hz, 3H), 0.97 (d, J=7.1Hz, 3H), 2.32-2.44 (m, 1H), 3.73-3.77 (m, 4H), 7.67 (q, J=3.3 Hz, 1H).

Compound 32b:(R)-2-((R)-2,2-Dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid methyl ester

Compound 32b was prepared in the same manner as compound 31b using 32ainstead of 31a in 30% yield. ¹H NMR (300 MHz, CDCl₃) δ0.91 (d, J=6.7 Hz,3H), 0.97 (d, J=6.7 Hz, 3H), 1.16 (s, 6H), 1.52-1.55 (m, 1H), 1.81-1.92(m, 1H), 2.76-2.85 (m, 1H), 3.20 (dd, J=9.8, 7.1 Hz, 1H), 3.72 (s, 3H),5.09 (d, J=17.4 Hz, 1H), 5.11 (d, J=10.8 Hz, 1H), 5.90 (dd, J=17.4, 10.8Hz, 1H). LCMS (m/z) 282.1 [M+H], Tr=3.51 min.

Compound 32c:(S)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid

A solution of 32b (1.30 g, 4.6 mmol) in tetrahydrofuran (30 mL) wasstirred at 0° C. under nitrogen. Potassium tert-butoxide (1.55 g, 13.8mmol) was added and the reaction mixture was stirred at 0° C. and warmedto room temperature overnight. The volatiles were evaporated, 2 Mhydrochloric acid was added and the mixture was extracted with ethylacetate. The organic extracts were combined, washed with water andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated to afford a mixture containing(R)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid tert-butyl ester,(S)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid tert-butyl ester,(R)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid and(S)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid (4.6 mmol) which was used crude in the next step. A solution of themixture of(R)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid tert-butyl ester,(S)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid tert-butyl ester,(R)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid and(S)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid (4.6 mmol) in dichloromethane (10 mL) was stirred at 5° C. undernitrogen. Trifluoroacetic acid (5 mL) was added and the reaction mixturewas stirred at room temperature for 20 h. Additional trifluoroaceticacid (5 mL) was added and the reaction mixture was stirred at roomtemperature for 48 h. The solvent was evaporated and the residue wasco-evaporated with toluene (3×). The residue was partitioned betweenethyl acetate and water. The organic extracts were combined, washed withwater and brine. The organics were filtered through a hydrophobic fritand the filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 10:1 to 5:1to afford(R)-2-((R)-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino)-3-methyl-butyricacid (152 mg, 12%) as a colorless oil and the title compound (321 mg,26%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ0.99 (d, J=6.7 Hz,3H), 1.01 (d, J=6.7 Hz, 3H), 1.23 (s, 6H), 2.09-2.19 (m, 1H), 2.85 (q,J=8.0 Hz, 1H), 3.27 (d, J=4.0 Hz, 1H), 5.16 (d, J=17.2 Hz, 1H), 5.19 (d,J=10.8 Hz, 1H), 5.91 (dd, J=17.3, 10.8 Hz, 1H). LCMS (m/z) 268.1 [M+H],Tr=2.95 min.

Compound 32d:(S)-2-{(E)-(R)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyricacid

Compound 32d was prepared in the same manner as compound 31d using 32cinstead of 31c and used crude in the next step. LCMS (m/z) 481.1 [M+H],Tr=3.15 min.

Compound 32e:(S)-1-[(S)-2-((S)-2-{(E)-(R)-4-[2-((R)-1-Acetoxy-ethyl)-quinolin-7-yl]-2,2-dimethyl-1-trifluoromethyl-but-3-enylamino}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

Compound 32e was prepared in the same manner as compound 31e using 32dinstead of 32c in 58% yield. ¹H NMR (300 MHz, CDCl₃) δ0.90 (d, J=7.1 Hz,3H), 0.92 (d, J=7.1 Hz, 3H), 1.31 (d, J=6.7 Hz, 3H), 1.38 (s, 6H), 1.69(d, J=6.7 Hz, 3H), 1.69-1.79 (m, 3H), 1.89-2.02 (m, 2H), 2.18 (s, 3H),2.18-2.21 (m, 1H), 2.83-3.12 (m, 3H), 3.66-3.73 (m, 1H), 3.83 (d, J=11.4Hz, 1H), 4.42-4.48 (m, 1H), 4.72 (d, J=11.8 Hz, 1H), 4.95 (d, J=11.8 Hz,1H), 5.27-5.34 (m, 1H), 6.06 (q, J=6.7 Hz, 1H), 6.56 (ABq, Δδ_(AB)=0.13,J_(AB)=16.3 Hz, 2H), 7.16 (d, J=7.6 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H),7.62 (d, J=8.5 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 8.02 (s, 1H), 8.12 (d,J=8.5 Hz, 1H). LCMS (m/z) 794.2/796.1 [M+H], Tr=3.65 min.

Compound 32

Compound 32 was prepared in the same manner as compound 31 using 32einstead of 31e in 35% as a white solid. ¹H NMR (300 MHz, CD₃CN) δ 0.94(d, J=6.9 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H), 1.19 (d, J=6.7 Hz, 3H), 1.43(s, 3H), 1.47 (s, 3H), 1.69 (d, J=6.7 Hz, 3H), 1.70-1.90 (m, 4H),2.18-2.22 (m, 2H), 3.39-3.60 (m, 3H), 3.81-3.96 (m, 2H), 4.73-4.81 (m,1H), 6.03-6.16 (m, 2H), 6.78 (ABq, Δδ_(AB)=0.10, J_(AB)=16.3 Hz, 2H),7.39 (d, J=8.5 Hz, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.87 (d, J=8.3 Hz, 1H),7.98-8.05 (br s, 1H), 8.25 (d, J=8.5 Hz, 1H), 8.34 (s, 1H). LCMS (m/z)604.3 [M+H], Tr=2.78 min.

Example 33 Compound 33

Compound 33a. (7-Bromo-quinolin-2-ylmethyl)-methyl-carbamic acidtert-butyl ester

To a stirred slurry of 7-bromo-quinoline-2-carbaldehyde (236 mg, 1.00mmol, prepared as in EP 239746) in methanol (5 mL) was added methylaminein methanol (2 M, 2.5 mL, 5.00 mmol). The reaction mixture was stirredfor 20 h and then evaporated. The residue was suspended in methanol (5mL) and sodium borohydride (57 mg, 1.50 mmol) was added portionwise. Thereaction mixture was stirred for 1 h and then partitioned betweendichloromethane and saturated ammonium chloride solution. The organiclayer was separated and washed with saturated ammonium chloride solutionand the aqueous extracts were extracted with dichloromethane and thecombined organics dried over anhydrous sodium sulfate, filtered andevaporated. The residue was dissolved in dichloromethane (8 mL) and tothe resulting stirred solution was added a solution of di-tert-butyldicarbonate (327 mg, 1.50 mmol) in dichloromethane (2 mL). Afterstirring for 3 h the reaction mixture was diluted with dichloromethaneand washed with saturated sodium bicarbonate solution (2×) and theaqueous layer was extracted with dichloromethane. The combined organicextracts were passed through a hydrophobic frit and evaporated. Theresidue was purified by silica gel chromatography eluting with agradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the titlecompound (206 mg, 59%) as an orange oil and as a mixture of rotamers. ¹HNMR (300 MHz, CDCl₃) δ1.42-1.61 (m, 9H), 2.92 (s, 1.5H), 3.00 (s, 1.5H),4.71 (s, 1H), 4.75 (s, 1H), 7.34-7.46 (m, 1H), 7.64 (d, J=8.5 Hz, 1H),7.70 (d, J=8.5 Hz, 1H), 8.13 (d, J=5.8 Hz, 1H), 8.26 (s, 1H). LCMS (m/z)351.1, 353.1 [M+H], Tr=3.13 min.

Compound 33b.(E)-4-{2-[(tert-Butoxycarbonyl-methyl-amino)-methyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid methyl ester

33a (206 mg, 0.586 mmol), 22d (171 mg, 0.674 mmol),bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)chloride (24 mg, 0.029 mmol) and potassium phosphate tribasic (373 mg,1.76 mmol) were suspended in cyclopentyl methyl ether (2 mL) and water(1 mL). The reaction mixture was stirred and heated at 80° C. for 2 hunder nitrogen. Morebis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)chloride (12 mg, 0.015 mmol) was added and the reaction mixture heatedat 90° C. for 90 minutes. The reaction was allowed to cool and dilutedwith ethyl acetate and washed with water (2×). The aqueous layers wereextracted with ethyl acetate and the combined organic extracts werewashed with saturated brine, dried over anhydrous sodium sulfate,filtered and evaporated. The residue was purified by silica gelchromatography eluting with a gradient of iso-hexanes/ethyl acetate 1:0to 7:3 to afford the title compound (178 mg, 76%) as a yellow gum and asa mixture of rotamers. ¹H NMR (300 MHz, CDCl₃) δ1.43-1.57 (m, 15H), 2.91(s, 1.5H), 2.99 (s, 1.5H), 3.74 (s, 3H), 4.71 (s, 1H), 4.74 (s, 1H),6.64 (app s, 2H), 7.27-7.39 (m, 1H), 7.63 (d, J=8.5 Hz, 1H), 7.76 (d,J=8.5 Hz, 1H), 7.90 (s, 1H), 8.10 (d, J=8.5 Hz, 1H). LCMS (m/z) 399.1[M+H], Tr=3.02 min.

Compound 33c.(E)-4-{2-[({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-methyl-amino)-methyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid methyl ester

33b (134 mg, 0.336 mmol) was suspended in hydrogen chloride in1,4-dioxane (4 M, 3.4 mL, 13.6 mmol) and the reaction mixture stirredfor 2.5 h and then evaporated. To the residue was added a solution of29a (106 mg, 0.353 mmol) in N,N-dimethylformamide (7 mL) followed byN,N-diisopropylethylamine (87 mg, 117 μL, 0.672 mmol),1-hydroxybenzotriazole hydrate (63 mg, 0.470 mmol) and N-(3dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (90 mg, 0.470mmol) and the reaction mixture was stirred for 16 h. The reactionmixture was diluted with ethyl acetate and washed successively withsaturated sodium bicarbonate solution, water, saturated ammoniumchloride solution, water and brine, dried over anhydrous sodium sulfate,filtered and evaporated. The residue was purified by silica gelchromatography eluting with a gradient of ethyl acetate/methanol 1:0 to19:1 to afford the title compound (128 mg, 65%) as a colorless gum. ¹HNMR (300 MHz, CDCl₃) δ0.80-1.06 (m, 6H), 1.21-1.38 (m, 3H), 1.47 (s,6H), 1.52-2.18 (m, 5H), 2.63-2.78 (m, 1H), 3.12 (s, 3H), 3.73 (s, 3H),3.88-3.97 (m, 2H), 4.08-4.26 (m, 2H), 4.41-4.61 (m, 1H), 4.75-4.99 (m,2H), 5.20-5.40 (m, 1H), 6.63 (app d, J=2.0 Hz, 2H), 7.22-7.34 (m, 1H),7.59-7.69 (m, 1H), 7.72-7.78 (m, 1H), 7.90-7.97 (d, J=8.0 Hz, 1H),8.08-8.20 (m, 1H). LCMS (m/z) 582.3 [M+H], Tr=2.35 min.

Compound 33

A stirred mixture of 33c (64 mg, 0.110 mmol) and 1M NaOH (0.88 mL, 0.88mmol) in 1,4-dioxane (5.5 mL) was heated at reflux for 2.5 hours andthen allowed to cool. The reaction mixture was neutralized to pH 7 with2M hydrochloric acid and evaporated. The residue was azeotroped withtert-butanol/acetonitrile/toluene and then acetonitrile/toluene to givethe crude acid.

To a stirred slurry of 2-methyl-6-nitrobenzoic acid (189 mg, 0.550mmol), 4-(dimethylamino)pyridine (101 mg, 0.825 mmol) and powdered 4 Åmolecular sieves (˜2 g) in 1,2-dichloroethane (37 mL) under nitrogen at50° C. was added a solution of the crude acid in N,N-dimethylformamide(5 mL) over 3 hours via syringe pump. The flask containing the originalacid was washed with further N,N-dimethylformamide (1 mL) and this wasadded to the reaction mixture over 10 minutes. The reaction mixture wasstirred at 50° C. for a further 2.5 hours and then allowed to cool. Themixture was filtered through celite and the filtrate washed successivelywith ice cold saturated ammonium chloride solution, ice cold saturatedsodium bicarbonate solution and saturated brine, dried over sodiumsulfate, filtered and evaporated. The residue was purified bypreparative HPLC using a gradient of acetonitrile/water 1:4 to 7:3 toafford the title compound (3.9 mg, 6.5%) as a white solid. LCMS (m/z)550.3 [M+H], Tr=2.35 min.

Examples 34 and 35 Compounds 34 and 35

Compound 34a.(S)-1-{(S)-2-[(S)-3-Methyl-2-(2-methyl-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

Compound 34a was prepared in the same manner as compound 27e using(S)-3-methyl-2-(2-methyl-but-3-enyloxy)-butyric acid instead of 27d in72% yield and isolated as a 1:1 mixture of diastereosiomers. LCMS (m/z)502.1/504.1 [M+H], Tr=3.32 min.

Compound 34b.(S)-1-{(S)-2-[(S)-3-Methyl-2-((E)-2-methyl-4-{2-[(R)-1-(2(R)-methyl-propane-2-sulfinylamino)-ethyl]-quinolin-7-yl}-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of 34a (642.0 mg, 1.282 mmol), 18c (455.4 mg, 1.282 mmol),N,N-dicyclohexylmethylamine (0.83 mL, 3.846 mmol) in 1,4-dioxane (10 mL)was heated to 100° C. for 2 h. After cooling to room temperature, thevolatiles were removed in vacuo and the residue was purified by silicagel chromatography using a 50 g Isolute cartridge eluted with acontinuous gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to affordthe title compound (500.1 mg, 50%) as an orange gum and as a 1:1 mixtureof diastereoisomers. LCMS (m/z) 774.3/776.2 [M+H], Tr=3.31 min.

Compound 34c.(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-2-methyl-4-{2-[(R)-1-((R)-2-methyl-propane-2-sulfinylamino)-ethyl]-quinolin-7-yl}-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid methyl ester

A cooled (0° C.) solution of 34b (500.1 mg, 0.645 mmol) intetrahydrofuran/water (30 mL, 5:1) was treated with lithium hydroxidemonohydrate (100.4 mg, 2.393 mmol). After stirring at 0° C. for 45minutes, the reaction was quenched with hydrogen chloride solution (2 M,1.2 mL) and the volatiles were removed in vacuo. The residualtrichloroethanol was azeotroped off with toluene, toluene/methanol (2×),toluene then dried in high vacuo. The white solid was triturated withdiethyl ether (2×) to provide crude(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-2-methyl-4-{2-[(R)-1-((R)-2-methyl-propane-2-sulfinylamino)-ethyl]-quinolin-7-yl}-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (LCMS (m/z) 644.4 [M+H], Tr=2.44 min) which was used withoutfurther purification. A cooled (0° C.) solution of crude(S)-1-{(S)-2-[(S)-3-methyl-2-((E)-2-methyl-4-{2-[(R)-1-((R)-2-methyl-propane-2-sulfinylamino)-ethyl]-quinolin-7-yl}-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (0.645 mmol), N,N-diisopropylethylamine (0.56 mL, 3.225 mmol) and4-dimethylaminopyridine (7.9 mg, 0.064 mmol) in dry tetrahydrofuran (210mL) was treated with 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate methanaminium (294.3 mg, 0.774 mmol). Thereaction was allowed to slowly warm to room temperature. After 16 h, thereaction was quenched with anhydrous methanol (5 mL). The volatiles wereremoved in vacuo and the residue was purified by silica gelchromatography using a 50 g Isolute cartridge eluted with a continuousgradient of dichloromethane/methanol 1:0 to 9:1 to afford the titlecompound (482.0 mg) as an orange gum and as a 1:1 mixture ofdiastereoisomers. LCMS (m/z) 658.3 [M+H], Tr=2.68 min.

Compound 34d:(S)-1-{(S)-2-[(S)-3-Methyl-2-((E)-2-methyl-4-{2-[(R)-1-((R)-2-methyl-propane-2-sulfinylamino)-ethyl]-quinolin-7-yl}-but-3-enyloxy)-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid

A solution of crude 34c (480 mg, 0.73 mmol) in tetrahydrofuran (15 mL)was stirred at 0° C. under nitrogen. A solution of lithium hydroxidemonohydrate (92 mg, 2.2 mmol) in water (3 mL) was added and the reactionmixture was stirred at 0° C. for 1 h. 1 M Hydrochloric acid (2.2 mL, 2.2mmol) was added and the volatiles were evaporated. The residue wasco-evaporated with tetrahydrofuran/toluene (1:1, 3×) and then trituratedwith diethyl ether (2×) and dried to afford the title compound (0.73mmol) as a pale yellow gum and as a 1:1 mixture of diastereoisomers.LCMS (m/z) 644.4 [M+H], Tr=2.44 min.

Compounds 34 and 35

A solution of crude 34d (500 mg, 0.5 mmol) in 1,4-dioxane (10 mL) wasstirred at room temperature under nitrogen. A solution of 4 M hydrogenchloride in 1,4-dioxane (1 mL) was added and the reaction mixture wasstirred at room temperature for 30 minutes. The volatiles wereevaporated and the residue was co-evaporated withtetrahydrofuran/toluene (1:1, 3×) and then triturated with diethyl ether(2×). The residue was dried to afford(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride as a dark yellow gum (0.5 mmol) and as a 1:1 mixtureof diastereoisomers which was used crude in the next reaction. LCMS(m/z) 540.2 [M+H], Tr=1.57 min. A suspension of(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-2-methyl-but-3-enyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.5 mmol) in dichloromethane (500 mL) was stirred at0° C. under nitrogen. N,N-Diisopropylethylamine (387 mg, 0.52 mL, 3.0mmol) and 4-dimethylaminopyridine (20 mg, 0.18 mmol) were added and thereaction mixture was stirred at 0° C. for 5 minutes.2-(1H-7-Azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (380 mg, 1.0 mmol) was added and thereaction mixture was stirred at 0° C. for 2 h and then at roomtemperature for 72 h. The volatiles were evaporated. Ethyl acetate wasadded and the mixture was washed with cold 1 M hydrochloric acid, water,saturated sodium hydrogen carbonate solution, water and brine. Theorganic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:3 toethyl acetate then ethyl acetate to ethyl acetate/methanol 1:10. Theresidue was purified by reverse phase preparative HPLC eluting withacetonitrile (containing 0.1% formic acid)/water (containing 0.1% formicacid) 35:65. Fractions containing the pure diastereoisomers werecombined and evaporated. The residues were partitioned between saturatedsodium hydrogen carbonate solution and ethyl acetate. The organic layerswere separated, washed with water and brine. The organic solutions werefiltered through a hydrophobic frit and the filtrate was dried to affordtwo diastereoisomers.

Compound 34 (First Eluting) Diastereomer 1: (3 mg, 1%) as a white solid.¹H NMR (300 MHz, CD₃OD) δ 0.97 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.7 Hz,3H), 1.25 (d, J=6.9 Hz, 3H), 1.59 (d, J=6.7 Hz, 3H), 1.60-1.65 (m, 2H),1.66 (d, J=7.1 Hz, 3H), 1.94-2.07 (m, 2H), 2.28-2.33 (m, 1H), 2.60-2.76(m, 2H), 3.49 (d, J=8.5 Hz, 1H), 3.55-3.71 (m, 2H), 3.92 (dd, J=9.5, 4.4Hz, 1H), 4.41-4.46 (m, 1H), 5.10 (q, J=6.5 Hz, 1H), 5.82 (q, J=7.1 Hz,1H), 6.51 (dd, J=16.1, 5.8 Hz, 1H), 6.63 (d, J=16.1 Hz, 1H), 7.43 (d,J=8.5 Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.82 (d, J=8.3 Hz, 1H), 8.02 (s,1H), 8.24 (d, J=8.5 Hz, 1H). LCMS (m/z) 522.2 [M+H], Tr=2.41 min.

Compound 35 (Second Eluting) Diastereomer 2: (3 mg, 1%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ 0.93 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.5Hz, 3H), 1.07 (d, J=6.9 Hz, 3H), 1.51-1.55 (m, 1H), 1.59 (d, J=6.5 Hz,3H), 1.68 (d, J=7.1 Hz, 3H), 1.69-1.73 (m, 1H), 1.94-1.99 (m, 2H),2.29-2.34 (m, 1H), 2.65-2.75 (m, 2H), 3.38-3.43 (m, 2H), 3.57-3.62 (m,1H), 3.96 (dd, J=9.0, 3.2 Hz, 1H), 4.40-4.46 (m, 1H), 5.10 (q, J=6.4 Hz,1H), 5.84-5.89 (m, 1H), 6.38 (dd, J=16.1, 6.3 Hz, 1H), 6.65 (d, J=16.1Hz, 1H), 7.42 (d, J=8.5 Hz, 1H), 7.46 (dd, J=8.5, 1.6 Hz, 1H), 7.82 (d,J=8.3 Hz, 1H), 8.05 (br s, 1H), 8.24 (d, J=8.5 Hz, 1H). LCMS (m/z) 522.2[M+H], Tr=2.38 min.

Example 36 Compound 36

Compound 36a.(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

Zinc dust (1.3 g, 20.3 mmol) was added to a solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (400 mg, 0.92 mmol) in tetrahydrofuran(20 mL). This suspension was treated with a solution of ammonium acetate(1.1 g, 13.9 mmol) in water (7 mL). After stirring at room temperaturefor 16 h, the zinc residues were filtered off and the volatiles wereremoved in vacuo. The residue was partitioned between ethyl acetate andsaturated potassium hydrogen sulfate solution, the aqueous layer wasextracted with ethyl acetate (2×), the organic layers were combined andthe volatiles were removed in vacuo. The residual acetic acid wasazeotroped off with toluene (3×10 mL) to give crude(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (280 mg, 99%) half of which (140 mg, 0.46 mmol) was combined with(R)-1-(7-bromo-quinolin-2-yl)-ethylamine hydrochloride (obtained fromASIBA Pharmatech, Inc.), (194 mg, 0.60 mmol) andN,N-diisopropylethylamine (0.400 mL, 2.3 mmol) in anhydrous acetonitrile(10 mL). This solution was then treated with2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (227 mg, 0.60 mmol). After stirring atroom temperature for 3 h, the volatiles were removed in vacuo. Theresidue was dissolved in ethyl acetate and subsequently washed withsaturated ammonium chloride solution and sodium bicarbonate solution.The organics were combined and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 25 g Biotagecartridge eluted with a continuous gradient of iso-hexanes/acetone 1:0to 1:1 to afford the title compound (136 mg, 55%) as a white solid. ¹HNMR (300 MHz, CD₃OD) δ 0.83 (d, J=6.9 Hz, 3H), 0.98 (d, J=6.9 Hz, 3H),1.38 (d, J=6.9 Hz, 3H), 1.60 (d, J=6.9 Hz, 3H), 1.65-1.75 (m, 3H),1.87-1.96 (m, 1H), 2.02-2.12 (m, 2H), 2.87-3.00 (m, 1H), 3.55-3.62 (m,1H), 3.81 (d, J=3.4 Hz, 1H), 4.24-4.34 (m, 1H), 5.18-5.28 (m, 1H),5.31-5.40 (m, 1H), 7.57 (d, J=8.7 Hz, 1H), 7.69 (dd, J=8.7, 1.8 Hz, 1H),7.85 (d, J=8.7 Hz, 1H), 8.26 (d, J=1.6 Hz, 1H), 8.33 (d, J=8.7 Hz, 1H).LCMS (m/z) 534.1, 536.1 [M+H], Tr=2.27 min.

Compound 36b. (R)-2-Methoxymethyl-2-methyl-but-3-enoic acid

Compound 36b was prepared in the same manner as compound 29d usingcompound(R)-4-isopropyl-3-((R)-2-methoxymethyl-2-methyl-but-3-enoyl)-oxazolidin-2-one(the enantiomer of 58a, prepared as described in Tet. Lett. 2000,41(33), 6429-6433) instead of 29c in 99% yield. ¹H NMR (300 MHz, CDCl₃)δ1.36 (s, 3H), 3.41 (s, 3H), 3.43 (d, J=8.9 Hz, 1H), 3.61 (d, J=8.9 Hz,1H), 5.20-5.30 (m, 2H), 6.00 (dd, J=17.6, 10.7 Hz, 1H).

Compound 36

A solution of 36a (123 mg, 0.23 mmol), 36b (40 mg, 0.27 mmol),palladium(II) acetate (11 mg, 0.046 mmol), tris-(o-tolyl)phosphine (21mg, 0.068 mmol) in anhydrous acetonitrile (3 mL) was treated withtriethylamine (81 μL, 0.57 mmol). After stirring at 100° C. undermicrowave irradiation for 20 minutes, the reaction was cooled to roomtemperature and the volatiles were removed in vacuo. The residue waspartitioned between dichloromethane and a saturated potassium hydrogensulfate solution. The organics were combined and the volatiles wereremoved in vacuo. The residue was azeotroped with toluene (3×10 mL) togive crude(E)-(R)-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-1-oxo-propyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2-methoxymethyl-2-methyl-but-3-enoicacid which was dissolved in N,N-dimethylformamide (5 mL) and added viasyringe pump over 6 h to a stirred slurry of 2-methyl-6-nitrobenzoicanhydride (161 mg, 0.47 mmol) and 4-dimethylaminopyridine (86 mg, 0.70mmol) in 1,2-dichloroethane (35 mL) at 50° C. under nitrogen. Thereaction was cooled down to room temperature and was subsequently washedwith 5% citric acid solution and sodium bicarbonate solution. Theorganics were combined and the volatiles were removed in vacuo. Theresidue was purified by preparative reverse phase HPLC to afford thetitle compound (8 mg, 15%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.01 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.7 Hz, 3H), 1.56 (s, 3H), 1.60 (d,J=6.9 Hz, 3H), 1.68 (d, J=6.9 Hz, 3H), 1.69-1.75 (m, 1H), 1.91-1.99 (m,1H), 2.10-2.30 (m, 2H), 3.36 (s, 2H), 3.41 (s, 3H), 3.49 (d, J=8.9 Hz,1H), 3.58-3.68 (m, 1H), 3.80 (d, J=9.1 Hz, 1H), 4.38-4.48 (m, 1H),5.03-5.13 (m, 1H), 5.19 (d, J=9.1 Hz, 1H), 5.74-5.85 (m, 1H), 6.41 (ABq,Δδ_(AB)=0.34, J_(AB)=16.4 Hz, 2H), 7.45 (d, J=8.5 Hz, 1H), 7.64 (d,J=8.5 Hz, 1H), 7.79 (s, 1H), 7.84 (d, J=8.5 Hz, 1H), 8.25 (d, J=8.5 Hz,1H). LCMS (m/z) 580.2 [M+H], Tr=2.38 min.

Example 37 Compound 37

Compound 37a. (7-Bromo-quinolin-2-ylmethyl)-(2,4-dimethoxy-benzyl)-amine

7-Bromo-quinoline-2-carbaldehyde (354 mg, 1.50 mmol, prepared asdescribed in EP 239746) was dissolved in 1,2-dichloroethane (15 mL) and2,4-dimethoxybenzylamine (251 mg, 225 μL, 1.50 mmol) was added and thereaction mixture stirred for 5 minutes before sodiumtriacetoxyborohydride (477 mg, 2.25 mmol) was added. After 3.5 hstirring the reaction mixture was quenched with saturated sodiumhydrogen carbonate solution and ethyl acetate was added. The organiclayer was separated and washed with sodium hydrogen carbonate and thecombined aqueous layers were extracted with ethyl acetate. The organicextracts were washed with saturated brine, dried over anhydrous sodiumsulfate, filtered and evaporated to give the crude, title compound as ayellow gum (571 mg) which was used without further purification. LCMS(m/z) 387.0, 389.0 [M+H], Tr=1.57 min.

Compound 37b. (7-Bromo-quinolin-2-ylmethyl)-carbamic acid tert-butylester

37a (290 mg, 0.75 mmol) was dissolved in dichloromethane (8 mL) andtriethylsilane (1.6 mL) and trifluoroacetic acid (8 mL) were added. Thereaction mixture was heated to reflux and stirred for 20 h and thenallowed to cool. The mixture was evaporated and purified by SCXcartridge eluting with methanol and then methanolic ammonia and thebasic fraction collected and evaporated. The residue was dissolved indichloromethane (8 mL) and di-tert-butyl dicarbonate (245 mg, 1.13 mmol)added and the reaction mixture stirred for 17 h. Diluted withdichloromethane and washed with saturated sodium hydrogen carbonatesolution (2×), the aqueous layer was back-extracted with dichloromethaneand the combined organic extracts passed through a hydrophobic frit andevaporated. The residue was purified by silica gel chromatography usinga gradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the titlecompound (150 mg, 59%) as a pale yellow solid. ¹H NMR (300 MHz, CDCl₃)δ1.52 (s, 9H), 4.65 (d, J=4.9 Hz, 2H), 5.90 (br s, 1H), 7.39 (d, J=8.5Hz, 2H), 7.63 (dd, J=8.5, 1.9 Hz, 2H), 7.72 (d, J=8.5 HZ, 1H), 8.11 (d,J=8.5 Hz, 1H), 8.28 (s, 1H). LCMS (m/z) 337.0, 339.0 [M+H], Tr=2.79 min.

Compound 37c.(E)-4-[2-(tert-Butoxycarbonylamino-methyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoicacid methyl ester

37b (296 mg, 0.878 mmol), 22d (256 mg, 1.01 mmol),bis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)chloride (36 mg, 0.044 mmol) and potassium phosphate tribasic (558 mg,2.63 mmol) were suspended in cyclopentyl methyl ether (3 mL) and water(1.5 mL). The reaction mixture was stirred and heated at 80° C. for 3 hunder nitrogen. The reaction was allowed to cool and diluted with ethylacetate and washed with water (2×) and brine, dried over anhydroussodium sulfate, filtered and evaporated. The residue was purified bysilica gel chromatography using a gradient of iso-hexanes/ethyl acetate1:0 to 7:3 to afford the title compound (257 mg, 76%) as a yellow solid.¹H NMR (300 MHz, CDCl₃) δ1.49 (s, 6H), 1.52 (s, 9H), 3.75 (s, 3H), 4.63(d, J=4.9 Hz, 2H), 5.96 (br s, 1H), 6.64 (s, 2H), 7.32 (d, J=8.5 Hz,1H), 7.63 (dd, J=8.5, 1.3 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 8.00 (s, 1H),8.08 (d, J=8.5 Hz, 1H). LCMS (m/z) 385.1 [M+H], Tr=2.64 min.

Compound 37d.(E)-4-{2-[({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-methyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid methyl ester

37c (128 mg, 0.333 mmol) was suspended in 4 M hydrogen chloride in1,4-dioxane (3.3 mL) and the reaction mixture stirred for 2.5 h and thenevaporated. To the residue was added a solution of 29a (105 mg, 0.350mmol) in N,N-dimethylformamide (7 mL) followed byN,N-diisopropylethylamine (86 mg, 116 μL, 0.666 mmol). To the stirredsolution was added 1-hydroxybenzotriazole hydrate (63 mg, 0.466 mmol)and N-(3 dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (89 mg,0.466 mmol) and the reaction mixture stirred for 16 h. The reactionmixture was diluted with ethyl acetate and washed successively withsaturated sodium bicarbonate solution, water, saturated ammoniumchloride solution, water and then brine, dried over anhydrous sodiumsulfate, filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of ethyl acetate/methanol 1:0 to 19:1 toafford the title compound (95 mg, 50%) as an off-white solid. ¹H NMR(300 MHz, CDCl₃) δ0.84 (d, J=6.9 Hz, 3H), 1.00 (d, J=6.9 Hz, 3H), 1.42(d, J=6.9 Hz, 3H), 1.48 (s, 6H), 1.65-1.76 (m, 2H), 1.88-1.98 (m, 1H),2.06-2.14 (m, 1H), 2.19-2.29 (m, 1H), 2.73-2.83 (m, 1H), 3.45-3.56 (m,1H), 3.73 (s, 3H), 3.95 (d, J=3.1 Hz, 1H), 3.99 (d, J=11.6 Hz, 1H),4.42-4.52 (m, 1H), 4.69-4.76 (m, 2H), 5.41-5.51 (m, 1H), 6.61-6.66 (m,1H), 6.74 (d, J=16.3 Hz, 1H), 7.24-7.31 (m, 1H), 7.62-7.69 (m, 1H),7.71-7.78 (m, 1H), 7.95-8.10 (m, 2H). LCMS (m/z) 568.3 [M+H], Tr=2.10min.

Compound 37e.(E)-4-{2-[({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-methyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid

A stirred mixture of 37d (95 mg, 0.167 mmol) and NaOH (1 M, 1.34 mL,1.34 mmol) in 1,4-dioxane (8 mL) was heated at reflux for 70 minutes andthen allowed to cool. The reaction mixture was acidified to pH 4 withhydrochloric acid (2 M) and evaporated. The residue was purified byreverse phase preparative HPLC using a gradient of acetonitrile/water1:19 to 7:3 to afford the title compound (43 mg, 47%) as a white solid.¹H NMR (300 MHz, CDCl₃) δ0.82 (d, J=6.8 Hz, 3H), 0.93-1.00 (m, 3H), 1.38(d, J=8.2 Hz, 3H), 1.49 (s, 6H), 1.58-1.76 (m, 2H), 1.81-1.94 (m, 1H),1.99-2.12 (m, 1H), 2.16-2.25 (m, 1H), 2.67-2.79 (m, 1H), 3.41-3.51 (m,1H), 3.92-4.00 (m, 1H), 4.01-4.08 (d, J=11.6 Hz, 1H), 4.40-4.54 (m, 1H),4.74 (d, J=4.5 Hz, 2H), 5.41-5.52 (m, 1H), 6.63-6.76 (m, 2H), 7.26-7.32(m, 1H), 7.33-7.39 (m, 1H), 7.55-7.64 (m, 1H), 7.75 (d, J=8.5 Hz, 1H),8.04-8.14 (m, 2H). LCMS (m/z) 554.2 [M+H], Tr=1.77 min.

Compound 37

To a stirred slurry of 2-methyl-6-nitrobenzoic acid (134 mg, 0.388mmol), 4-(dimethylamino)pyridine (71 mg, 0.583 mmol) and powdered 4 Åmolecular sieves (˜1 g) in 1,2-dichloroethane (26 mL) under nitrogen at50° C. was added a solution of 37e (43 mg, 0.078 mmol) inN,N-dimethylformamide (3 mL) over 4 h via syringe pump. The flaskcontaining the original acid was washed with furtherN,N-dimethylformamide (1 mL) and this was added to the reaction mixtureover 10 minutes. After the end of addition, the reaction mixture wasstirred at 50° C. for 3 h and then allowed to cool. The mixture wasfiltered through Celite and the filtrate washed successively withice-cold saturated ammonium chloride solution, ice-cold saturated sodiumbicarbonate solution and saturated brine, dried over anhydrous sodiumsulfate, filtered and evaporated. The residue was purified by reversephase preparative HPLC using a gradient of acetonitrile/water 1:4 to 4:1modified with 0.1% formic acid and the product triturated with ether toafford the title compound (6.4 mg, 15%) as a white solid. ¹H NMR (300MHz, CD₃OD) δ 1.02 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.9 Hz, 3H), 1.46 (s,3H), 1.48 (d, J=7.3 Hz, 3H), 1.54 (s, 3H), 1.55-1.79 (m, 2H), 1.94-2.04(m, 1H), 2.16-2.30 (m, 1H), 2.30-2.40 (m, 1H), 2.71-2.83 (m, 1H),3.53-3.60 (m, 1H), 4.42-4.51 (m, 1H), 4.67, 4.75 (ABq, J_(AB)=18.4 Hz,2H), 5.20 (d, J=8.9 Hz, 2H), 5.89 (q, J=7.2, 1H), 6.44 (d, J=16.2 Hz,1H), 6.69 (d, J=16.2 Hz, 1H), 7.40 (d, J=8.5 Hz, 1H), 7.56 (d, J=8.5,1.3 Hz, 1H), 7.85 (d, J=8.5 Hz, 1H), 7.92 (s, 1H), 8.23 (d, J=8.5 Hz,1H). LCMS (m/z) 536.2 [M+H], Tr=2.47 min.

Example 38 Compound 38

Compound 38a (R)-2-Methyl-propane-2-sulfinic acid[1-(7-bromo-quinolin-2-yl)-eth-(E)-ylidene]-amide

To a solution of 18a (1.42 g, 5.68 mmol) in THF (28 mL) was addedtitanium (IV) ethoxide (2.6 g, 2.35 mL, 11.4 mmol, tech. grade) followedby (R)-(+)-2-methyl-propanesulfinimide (825 mg, 6.82 mmol). The reactionmixture was stirred at 60° C. under nitrogen for 6 hours and allowed tocool. Brine was added followed by ethyl acetate and the suspensionfiltered through celite and the filter pad was washed with ethylacetate. The ethyl acetate layer was separated, dried over sodiumsulfate, filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexane/ethyl acetate 9:1 to 3:1to give the title compound (448 mg, 22%) as an orange solid. ¹H NMR (300MHz, CDCl₃) δ 1.38 (s, 9H), 2.99 (s, 3H), 7.71 (m, 2H), 8.16 (d, J=8.6Hz, 1H), 8.24 (d, J=8.6 Hz, 1H), 8.37 (s, 1H). LCMS (m/z) 352.9/354.9[M+H], Tr 3.14 minutes.

Compound 38b (R)-2-Methyl-propane-2-sulfinic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

A mixture of (1S,2R)-(−)-cis-1-amino-2-indanol (19 mg, 0.13 mmol), [Ru(p-cymen) Cl₂]₂ (39 mg, 0.064 mmol) and powdered 4 Å Molecular Sieves(0.7 g) was suspended in anhydrous 2-propanol (3 mL) and stirred undernitrogen. The suspension was heated at 90° C. for 30 minutes. Thereaction mixture was cooled to 40° C. and a solution 38a (448 mg, 1.27mmol) in 2-propanol (9 mL) was added followed by a solution of potassiumtert-butoxide (36 mg, 0.32 mmol) in 2-propanol (3 mL). The reactionmixture was stirred for 2 hours at 40° C. and then allowed to cool. Themixture was poured directly onto a silica gel cartridge and eluted withethyl acetate. After concentration the residue was further purified bysilica gel chromatography using a gradient of iso-hexane/ethyl acetate1:1 to 0:1 to give the title compound (287 mg, 64%) as a brown gum. ¹HNMR (300 MHz, CDCl₃) δ 1.33 (s, 9H), 1.60 (d, J=6.7 Hz, 3H), 4.80 (m,1H), 5.42 (br d, J=4.2 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H), 7.62 (dd, J=8.5,1.2 Hz, 1H), 7.68 (d, J=8.7 Hz, 1H), 8.12 (d, J=8.7 Hz, 1H), 8.25 (s,1H). LCMS (m/z) 354.9/356.8 [M+H], Tr 2.49 min

Compound 38c. 2-Methyl-propane-2-sulfinic acid(R)-[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-methyl-amide

A cooled (−20° C.) solution of 38b (100 mg, 0.28 mmol, same as 18c) inN,N-dimethylformamide (5 mL) was treated with lithiumbis(trimethylsilyl)amide (0.28 mL, 0.28 mmol, 1 M in hexane). Afterstirring at this temperature for 1 h, iodomethane (0.035 mL, 0.56 mmol)was added and the temperature raised to room temperature. After stirringat room temperature for 1 h, the reaction was quenched by addition ofwater (10 mL). The mixture was extracted with diethyl ether (3×20 mL).The organics were combined and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 10 g Biotagecartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate1:0 to 1:1 to afford the title compound (149 mg, 62%) as a white solid.¹H NMR (300 MHz, CDCl₃) δ1.25 (s, 9H), 1.74 (d, J=6.9 Hz, 3H), 2.53 (s,3H), 4.81 (q, J=6.9 Hz, 1H), 7.55-7.70 (m, 3H), 8.11 (d, J=8.5 Hz, 1H),8.27 (s, 1H). LCMS (m/z) 369.0, 371.0 [M+H], Tr=2.91 min.

Compound 38d.(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-methyl-amide

A solution of 38c (450 mg, 1.22 mmol) in methanol (10 mL) was treatedwith hydrochloric acid in 1,4-dioxane (4 M, 5 mL). After stirring atroom temperature for 2 h, the volatiles were removed in vacuo. Theresidual solvent was azeotroped off with toluene (3×10 mL) to give crude[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-methyl-amine hydrochloride.

Zinc dust (1.30 g, 20.32 mmol) was added to a solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (400 mg, 0.92 mmol) in tetrahydrofuran(20 mL). This suspension was treated with a solution of ammonium acetate(1.10 g, 13.9 mmol) in water (7 mL). After stirring at room temperaturefor 16 h, zinc residues were filtered off and the volatiles were removedin vacuo. The residue was partitioned between ethyl acetate andsaturated potassium hydrogen sulfate solution, the aqueous layer wasextracted with ethyl acetate (2×), the organic layers were combined andthe volatiles were removed in vacuo. The residual acetic acid wasazeotroped off with toluene (3×10 mL) to give crude(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid which was combined with crude[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-methyl-amine hydrochloride inanhydrous acetonitrile (15 mL) and N,N-diisopropylethylamine (0.801 mL,4.60 mmol). This solution was then treated with2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (464 mg, 1.22 mmol). After stirring atroom temperature for 2 h, the volatiles were removed in vacuo. Theresidue was dissolved in ethyl acetate and subsequently washed withsaturated ammonium chloride solution and sodium bicarbonate solution.The organics were combined and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 25 g Biotagecartridge eluted with a continuous gradient of iso-hexanes/acetone 1:0to 0:1 to afford the title compound (318 mg, 58%) as a white solid. ¹HNMR (300 MHz, CD₃OD) δ0.85 (d, J=6.9 Hz, 3H), 1.00 (d, J=6.9 Hz, 3H),1.33 (d, J=6.9 Hz, 3H), 1.69 (d, J=6.9 Hz, 3H), 1.72-2.15 (m, 6H), 2.98(s, 3H), 2.85-2.95 (m, 1H), 3.80-3.90 (m, 1H), 4.30-4.40 (m, 1H),5.25-5.35 (m, 1H), 5.95-6.10 (m, 1H), 7.45-7.78 (m, 1H), 7.65-7.72 (m,1H), 7.80-7.90 (m, 1H), 8.18-8-38 (m, 2H). LCMS (m/z) 548.1, 550.1[M+H], Tr=2.60 min.

Compound 38

Compound 38 was prepared in the same manner as compound 36 using 38dinstead of (36a (123 mg, 0.23 mmol) in 4% yield. ¹H NMR (300 MHz, CD₃OD)δ1.00 (d, J=6.7 Hz, 3H), 1.01 (d, J=6.7 Hz, 3H), 1.52-1.59 (m, 6H), 1.73(d, J=7.4 Hz, 3H), 1.76-1.88 (m, 2H), 1.89-1.99 (m, 2H), 2.09-2.21 (m,1H), 2.71-2.84 (m, 1H), 3.29 (s, 3H), 3.40 (s, 3H), 3.44 (d, J=8.7 Hz,1H), 3.79 (d, J=8.9 Hz, 1H), 4.13-4.22 (m, 1H), 4.39-4.47 (m, 1H), 5.05(d, J=9.1 Hz, 1H), 5.83-5.97 (m, 2H), 6.38 (ABq, Δδ_(AB)=0.24,J_(AB)=16.5 Hz, 2H), 7.45 (d, J=8.5 Hz, 1H), 7.64-7.73 (m, 2H), 7.79 (d,J=8.5 Hz, 1H), 8.19 (d, J=8.5 Hz, 1H). LCMS (m/z) 594.2 [M+H], Tr=2.48min.

Examples 39 and 40 Compound 39 and 40

Compound 39a. [(R)-1-(7-Bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester

To a solution of (R)-1-(7-bromo-quinolin-2-yl)-ethylamine hydrochloride(obtained from ASIBA Pharmatech Inc.), (3.5 g, 12.2 mmol) in anhydrousdichloromethane at 0° C. and under an atmosphere of nitrogen was addedtriethylamine (5.1 mL, 36.5 mmol), 4-dimethylaminopyridine (297 mg, 2.4mmol) and di-tert-butyl dicarbonate (4.0 g, 18.3 mmol). The reaction waswarmed to room temperature and stirred for 2 h. The reaction was cooledto 0° C. and quenched with hydrochloric acid (1 M). The organic layerwas separated, washed with brine, dried through a hydrophobic frit andconcentrated in vacuo. The product was purified by silica gelchromatography using iso-hexanes/ethyl acetate 6:1 to afford the titlecompound (3 g, 70%) as a yellow solid. ¹H NMR (300 MHz, CDCl₃) δ1.49 (s,9H), 1.55 (d, J=6.9 Hz, 3H), 4.93-5.10 (m, 1H), 6.01-6.20 (m, 1H), 7.38(d, J=8.5 Hz, 1H), 7.62 (dd, J=8.7, 1.8 Hz, 1H), 7.69 (d, J=8.7 Hz, 1H),8.11 (d, J=8.5 Hz, 1H), 8.29 (s, 1H). LCMS (m/z) 353.0 [M+H], Tr=3.02min.

Compound 39b.(E)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoicacid methyl ester

To a solution 39a (Ig, 2.86 mmol) in cyclopentylmethyl ether (14 mL) andwater (7 mL), at room temperature, was added 22d (840 mg, 3.43 mmol),potassium phosphate tribasic (1.8 g, 8.57 mmol) andbis[(dicyclohexyl)(4-dimethylaminophenyl)phosphine]palladium(II)chloride (116 mg, 0.14 mmol). The reaction was heated to 80° C. for 2 h.The reaction was cooled, diluted with ethyl acetate and washed withbrine (2×). The organic layer was dried through a hydrophobic frit andconcentrated in vacuo. The product was purified by silica gelchromatography using iso-hexanes/ethyl acetate 2:1 to afford the titlecompound (1.1 g, 96%) as a viscous oil. ¹H NMR (300 MHz, CDCl₃)δ1.43-1.52 (m, 15H), 1.56 (d, J=6.7 Hz, 3H), 3.75 (s, 3H), 4.92-5.09 (m,1H), 6.16-6.32 (m, 1H), 6.64 (s, 2H), 7.30 (d, J=8.5 Hz, 1H), 7.62 (dd,J=8.5, 1.6 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 8.02 (s, 1H), 8.08 (d, J=8.5Hz, 1H). LCMS (m/z) 399.3 [M+H], Tr=2.75 min.

Compound 39c.(E)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoicacid

To a solution of 39b (1.1 g, 2.76 mmol) in tetrahydrofuran (24 mL) andwater (5 mL) was added lithium hydroxide monohydrate (580 mg, 13.8mmol). The reaction was heated to 40° C. for 20 h. The reaction wascooled to room temperature and acidified to pH 5 with aqueoushydrochloric acid (2 M). The mixture was evaporated to dryness anddichloromethane and acetonitrile added. This suspension was filteredthrough a hydrophobic frit and the filtrate concentrated in vacuo toafford the title compound (800 mg, 76%) as a yellow solid. ¹H NMR (300MHz, CDCl₃) δ1.46 (s, 9H), 1.51 (s, 6H), 1.56 (d, J=6.7 Hz, 3H),4.91-5.14 (m, 1H), 6.35-6.52 (m, 1H), 6.66, 6.72 (ABq, J=16.2 Hz, 2H),7.31 (d, J=8.5 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.70 (d, J=8.3 Hz, 1H),7.99-8.17 (m, 2H). LCMS (m/z) 385.3 [M+H], Tr=2.21 min.

Compound 39d:(S)-1-(2-tert-Butoxycarbonylamino-3-methoxy-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

Compound 39d was prepared (starting from bis-N-Boc TCE ester prepared asdescribed in Angew. Chem. Intl. Ed. English, 1999, 38, 2443.) in thesame manner as intermediate 1d using N-α-Boc-O-methyl serine (1 g, 4.56mmol) instead of L-N-Boc-alanine to afford the title compound as a clearviscous oil (1.5 g, 71%) and as a 1:1 mixture of two diastereoisomers.LCMS (m/z) 462.7 [M+H], Tr=2.71 min.

Compound 39e.(S)-1-[2-((S)-2-Hydroxy-3-methyl-butyrylamino)-3-methoxy-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

To a solution of 39d (1.0 g, 2.16 mmol) in anhydrous dichloromethane (7mL) at room temperature and under an atmosphere of nitrogen was addedtrifluoroacetic acid (1.7 mL). The reaction was stirred at roomtemperature for 1.5 h and concentrated in vacuo. The ensuing residue wasco-evaporated from toluene (3×) to yield a viscous light brown oil. Thiswas dissolved in anhydrous acetonitrile (22 mL) and cooled to 0° C.before adding (S)-2-hydroxy-3-methyl-butyric acid (255 mg, 2.2 mmol),hydroxybenzotriazole monohydrate (497 mg, 3.3 mmol),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (623 mg, 3.3 mmol) andN,N-diisopropylethylamine (1.9 mL, 10.8 mmol). The reaction mixture waswarmed to ambient temperature and stirred for 72 h. The reaction wasdiluted with ethyl acetate and washed with hydrochloric acid (1 M),saturated sodium bicarbonate and brine. The organic layer was driedthrough a hydrophobic frit and concentrated in vacuo. The residue waspurified by silica gel chromatography using iso-hexanes/acetone 3:2 togive the title compound (750 mg, 75% over 2 steps) as a clear viscousoil and as a 1:1 mixture of two diastereoisomers. LCMS (m/z) 462.0[M+H], Tr=2.24 min.

Compound 39f.(S)-1-[2-((S)-2-{(E)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-3-methoxy-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro ethyl ester

To a solution of 39b (300 mg, 0.78 mmol) in anhydrous dichloromethane (5mL) at room temperature and under an atmosphere of nitrogen was addedN,N-diisopropylethylamine (0.28 mL, 1.56 mmol), 4-dimethylaminopyridine(200 mg, 1.64 mmol) and 2-methyl-6-nitrobenzoic anhydride (323 mg, 0.94mmol) followed by a solution of 39e (433 mg, 0.94 mmol) in anhydrousdichloromethane (2.5 mL). The reaction was stirred for 2 h after whichit was diluted with ethyl acetate and washed with water and brine. Theorganic layer was dried through a hydrophobic frit and concentrated invacuo. The residue was purified by silica gel chromatography usingiso-hexanes/ethyl acetate 1:2 to give the title compound (211 mg, 33%)as a clear viscous oil and as a 1:1 mixture of two diastereoisomers.LCMS (m/z) 828.3 [M+H], Tr=3.48 min.

Compounds 39 and 40

To a solution of 39f (200 mg, 0.24 mmol) in anhydrous tetrahydrofuran (3mL) at 0° C. was added a 0.4 M aqueous solution of sodium hydroxide (0.7mL, 0.29 mmol). The reaction was stirred at 0° C. for 1 h before beingacidified to pH 5 with hydrochloric acid (2 M) and concentrated invacuo. The resulting residue was partitioned between dichloromethane andwater and the organic layer separated, dried through a hydrophobic fritand concentrated in vacuo. The residue was dissolved in anhydrous1,4-dioxane (1 mL) and at room temperature was added a 4 M solution ofhydrochloric acid in dioxane (0.3 mL, 1.2 mmol). The reaction wasstirred for 1.5 h, concentrated in vacuo and the resulting solidtriturated with diethyl ether to afford a light yellow solid. The solidwas dissolved in anhydrous N,N-dimethylformamide (3 mL) andN,N-diisopropylethylamine (0.21 mL, 1.2 mmol) and this solution wasadded to a pre-stirred solution of2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (137 mg, 0.4 mmol) in anhydrousdichloromethane (80 mL) at 0° C. and under an atmosphere of nitrogen.Following the addition, the reaction was stirred at room temperature for2 h before being concentrated in vacuo. The ensuing residue wasdissolved in ethyl acetate and washed with hydrochloric acid (0.5 M),saturated sodium bicarbonate and brine. The organic layer was driedthrough a hydrophobic frit and concentrated in vacuo. The residue waspurified by reverse phase preparatory HPLC to afford twodiastereoisomers as white solids.

Compound 39. (Second eluting) Diastereoisomer 1 (7 mg, 5%). ¹H NMR (300MHz, CD₃OD) δ1.06 (d, J=6.7 Hz, 3H), 1.09 (d, J=6.7 Hz, 3H), 1.48 (s,3H), 1.57 (d, J=6.7 Hz, 3H), 1.59 (s, 3H), 1.51-1.65 (m, 1H), 1.69-2.11(m, 4H), 2.19-2.37 (m, 1H), 2.72-2.86 (m, 1H), 2.91 (s, 3H), 3.40-3.58(m, 3H), 3.80-3.90 (m, 1H), 4.33-4.45 (m, 1H), 5.11-5.22 (m, 3H),5.57-5.63 (m, 1H), 6.54, 6.71 (ABq, J=15.9 Hz, 2H), 7.44 (d, J=8.5 Hz,1H), 7.54 (d, J=8.5 Hz, 1H), 7.85 (d, J=8.5 Hz, 1H), 7.96 (s, 1H), 8.24(d, J=8.5 Hz, 1H). LCMS (m/z) 580.2 [M+H], Tr=2.58 min.

Compound 40 (First eluting) Diastereomer 2. (3 mg, 2%); ¹H NMR (300 MHz,CD₃OD) δ 1.00 (d, J=6.7 Hz, 3H), 1.05 (d, J=6.7 Hz, 3H), 1.43 (s, 3H),1.54 (s, 3H), 1.57 (d, J=6.7 Hz, 3H), 1.53-1.75 (m, 1H), 1.91-2.02 (m,1H), 2.12-2.24 (m, 1H), 2.25-2.36 (m, 1H), 2.64-2.76 (m, 1H), 3.46 (s,3H), 3.59-3.70 (m, 1H), 3.87 (d, J=6.0 Hz, 2H), 4.41-4.50 (m, 1H), 5.12(q, J=6.7 Hz, 1H), 5.18 (d, J=11.6 Hz, 1H), 5.27 (d, J=8.5 Hz, 1H), 6.09(t, J=6.3 Hz, 1H), 6.41, 6.61 (ABq, J=16.3 Hz, 2H), 7.43 (d, J=8.5 Hz,1H), 7.58 (dd, J=8.5, 1.5 Hz, 1H), 7.83 (d, J=8.5 Hz, 1H), 8.03 (s, 1H),8.24 (d, J=8.5 Hz, 1H). LCMS (m/z) 580.2 [M+H], Tr=2.62 min.

Example 41 Compound 41

Compound 41a. [(R)-1-(7-Bromo-quinolin-2-yl)-ethyl]-methyl-carbamic acidtert-butyl ester

To a solution 39a (500 mg, 1.4 mmol) in anhydrous N,N-dimethylformamide(5 mL), at −20° C. and under an atmosphere of nitrogen, was addeddropwise lithium bis(trimethylsilyl)amide (1.0 M in THF, 1.7 mL, 1.71mmol) over 10 minutes. Following the addition the reaction was stirredat −20° C. for 45 minutes and then iodomethane (0.18 mL, 2.85 mmol) wasadded. The reaction was warmed to room temperature and stirred for 1 hbefore quenching with saturated aqueous ammonium chloride and extractingwith diethyl ether (2×). The combined organics were washed with 5%aqueous lithium chloride solution, dried through a hydrophobic frit andconcentrated in vacuo. The product was purified by silica gelchromatography using iso-hexanes/ethyl acetate 6:1 to afford the titlecompound (500 mg, 96%) as a viscous oil. ¹H NMR (300 MHz, CDCl₃) δ1.50(s, 9H), 1.68 (d, J=6.9 Hz, 3H), 2.72 (br s, 3H), 5.34-5.92 (m, 1H),7.41 (d, J=8.5 Hz, 1H), 7.62 (dd, J=8.5, 1.8 Hz, 1H), 7.68 (d, J=8.7 Hz,1H), 8.08 (d, J=8.7 Hz, 1H), 8.29 (d, J=1.6 Hz, 1H). LCMS (m/z) 367.0[M+H], Tr=3.61 min.

Compound 41b.(E)-4-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid methyl ester

Compound 41b was prepared in the same manner as compound 39b using 41ainstead of 39a in 80% yield. ¹H NMR (300 MHz, CDCl₃) δ1.50 (s, 9H), 1.56(s, 6H), 1.69 (d, J=7.1 Hz, 3H), 2.72 (br s, 3H), 3.74 (s, 3H),5.28-6.01 (m, 1H), 6.64 (s, 2H), 7.33 (d, J=8.3 Hz, 1H), 7.61 (dd,J=8.7, 1.6 Hz, 1H), 7.74 (d, J=8.3 Hz, 1H), 8.01-8.08 (m, 2H). LCMS(m/z) 413.2 [M+H], Tr=3.42 min.

Compound 41c.(E)-4-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid

Compound 41c was prepared in the same manner as compound 39c using 41binstead of(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoicacid methyl ester in 99% yield. ¹H NMR (300 MHz, CDCl₃) δ1.49 (s, 9H),1.52 (s, 6H), 1.69 (d, J=7.1 Hz, 3H), 2.73 (br s, 3H), 5.29-5.84 (m,1H), 6.66, 6.70 (ABq, J=16.3 Hz, 2H), 7.34 (d, J=108.9 Hz, 1H),7.59-7.66 (m, 1H), 7.74 (d, J=8.5 Hz, 1H), 8.00-8.09 (m, 2H). LCMS (m/z)399.1 [M+H], Tr=2.75 min.

Compound 41d.(S)-1-[(S)-2-tert-Butoxycarbonylamino-3-(tert-butyl-diphenyl-silanyloxy)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

To (S)-tetrahydro-pyridazine-1,2,3-tricarboxylic acid 1,2-di-tert-butylester 3-(2,2,2-trichloro-ethyl) ester (prepared according to theprocedure described in Angew. Chem. Int. Ed. English 1999, 38, 2443.),(5.0 g, 10.8 mmol) in anhydrous dichloromethane (33 mL) at 0° C. andunder an atmosphere of nitrogen was added trifluoroacetic acid (33 mL,432 mmol). The solution was warmed to room temperature and stirred for16 h. The reaction was concentrated in vacuo and the residueco-evaporated from toluene (3×). The resulting brown viscous oil wasdissolved in anhydrous acetonitrile (5 mL) and added to a solution of(S)-2-tert-butoxycarbonylamino-3-(tert-butyl-diphenyl-silanyloxy)-propionicacid (2.39 g, 5.4 mmol, prepared as in PCT Int. Appl. 2006, WO2006004880 A2), N,N-diisopropylethylamine (3.76 mL, 21.6 mmol) andO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(2.05 g, 5.4 mmol) in anhydrous acetonitrile (25 mL) that had beenpreviously stirred at 0° C. for 20 minutes. The reaction was warmed toroom temperature and stirred for 16 h before being concentrated invacuo. The residue was purified by silica gel chromatography usingiso-hexanes/ethyl acetate 4:1 to give the title compound (1.8 g, 49%) asa clear viscous oil. ¹H NMR (300 MHz, CDCl₃) δ1.08 (s, 9H), 1.46 (s,9H), 1.53-1.67 (m, 1H), 1.80-2.01 (m, 2H), 2.82-2.96 (m, 1H), 3.19-3.32(m, 1H), 3.62 (d, J=10.9 Hz, 1H), 3.80-3.97 (m, 3H), 4.23-4.34 (m, 1H),4.61 (d, J=12.0 Hz, 1H), 4.90 (d, J=12.0 Hz, 1H), 5.18-5.27 (m, 1H),5.51-5.63 (m, 1H), 7.35-7.49 (m, 6H), 7.58-7.70 (m, 4H). LCMS (m/z)686.2 [M+H], Tr=4.32 min.

Compound 41e.(S)-1-[(S)-3-(tert-Butyl-diphenyl-silanyloxy)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

Compound 41e was prepared in the same manner as compound 39e using(S)-1-[(S)-2-tert-butoxycarbonylamino-3-(tert-butyl-diphenyl-silanyloxy)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester instead of(S)-1-((S)-2-tert-butoxycarbonylamino-3-methoxy-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester in 50% yield. ¹H NMR (300 MHz, CDCl₃)δ0.90 (d, J=6.7 Hz, 3H), 1.07-1.09 (m, 12H), 1.42-1.71 (m, 2H),1.80-1.91 (m, 1H), 1.93-2.02 (m, 1H), 2.07-2.15 (m, 1H), 2.72 (d, J=5.9Hz, 1H), 2.86-3.02 (m, 1H), 3.17-3.28 (m, 1H), 3.60 (d, J=10.5 Hz, 1H),3.90-3.99 (m, 3H), 4.18-4.29 (m, 1H), 4.62, 4.91 (ABq, J=11.9 Hz, 2H),5.42-5.50 (m, 1H), 7.35-7.51 (m, 7H) 7.58-7.71 (m, 4H). LCMS (m/z) 686.2[M+H], Tr=3.93 min.

Compound 41f

Compound 41f was prepared in the same manner as compound 39f using(E)-4-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid and(S)-1-[(S)-3-(tert-butyl-diphenyl-silanyloxy)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester instead of(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoicacid and(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-3-methoxy-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester in 32% yield. ¹H NMR (300 MHz, CDCl₃)δ0.98-1.05 (m, 6H), 1.07 (s, 9H), 1.50 (s, 9H), 1.52-1.60 (m, 2H), 1.57(s, 3H), 1.61 (s, 3H), 1.68 (d, J=6.9 Hz, 3H), 1.72-1.93 (m, 2H),2.31-2.46 (m, 1H), 2.63-3.01 (m, 5H), 3.47 (d, J=11.2 Hz, 1H), 3.87 (dd,J=10.3, 2.7 Hz, 1H), 4.04 (dd, J=10.3, 3.4 Hz, 1H), 4.11-4.23 (m, 1H),4.60, 4.88 (ABq, J=12.0 Hz, 2H), 5.21 (d, J=3.6 Hz, 1H), 5.38-5.44 (m,1H), 6.72, 6.76 (ABq, J=16.3 Hz, 2H), 7.31-7.49 (m, 8H), 7.56-7.62 (m,2H), 7.64-7.71 (m, 4H), 7.99-8.06 (m, 2H). LCMS (m/z) 1066.5 [M+H],Tr=4.94 min.

Compound 41g

Compound 41g was prepared in the same manner as compound 39 using 41finstead of(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-dimethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-3-methoxy-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro ethyl ester in 77% yield. ¹H NMR (300 MHz, CDCl₃)δ0.95-1.04 (m, 6H), 1.46 (s, 3H), 1.53 (s, 3H), 1.54-1.61 (m, 12H),1.65-2.01 (m, 4H), 2.19-2.37 (m, 1H), 2.61 (s, 3H), 2.65-2.78 (m, 1H),3.57-3.76 (m, 2H), 3.79 (d, J=12.0 Hz, 1H), 4.03-4.10 (m, 1H), 4.52-4.63(m, 1H), 4.89 (d, J=8.5 Hz, 1H), 5.93 (q, J=7.1 Hz, 1H), 5.98-6.07 (m,1H), 6.28, 6.47 (ABq, J=16.4 Hz, 2H), 6.56-6.63 (m, 1H), 7.21-7.54 (m,7H), 7.61-7.75 (m, 7H), 8.02 (d, J=8.3 Hz, 1H). LCMS (m/z) 818.4 [M+H],Tr=4.22 min.

Compound 41

To a solution of compound 41g (190 mg, 0.23 mmol) in anhydroustetrahydrofuran (1 mL), at room temperature and under an atmosphere ofnitrogen, was added tetrabutylammonium fluoride (1 M in THF, 1.2 mL, 1.2mmol). The reaction was stirred for 45 minutes before being diluted withethyl acetate and washed with saturated aqueous ammonium chloride. Theorganic layer was dried through a hydrophobic frit and concentrated invacuo. The product was purified by silica gel chromatography usingiso-hexanes/acetone 1:1 to give the title compound (35 mg, 27%) as awhite solid. ¹H NMR (300 MHz, CD₃OD) δ0.98 (d, J=6.7 Hz, 3H), 1.01 (d,J=6.5 Hz, 3H), 1.42 (s, 3H), 1.52 (s, 3H), 1.72 (d, J=7.4 Hz, 3H),1.72-1.97 (m, 4H), 2.09-2.25 (m, 1H), 2.71-2.86 (m, 1H), 3.30 (s, 3H),3.96 (d, J=6.3 Hz, 2H), 4.18-4.30 (m, 1H), 4.42-4.53 (m, 2H), 5.08 (d,J=8.7 Hz, 1H), 5.88 (q, J=7.4 Hz, 1H), 6.06 (t, J=6.0 Hz, 1H), 6.40,6.49 (ABq, J=16.5 Hz, 2H), 7.44 (d, J=8.7 Hz, 1H), 7.64 (dd, J=8.5, 1.3Hz, 1H), 7.78 (d, J=8.5 Hz, 1H), 7.89 (s, 1H), 8.19 (d, J=8.7 Hz, 1H).LCMS (m/z) 580.2 [M+H], Tr=2.14 min.

Example 42 Compound 42

Compound 42a. 4-Oxo-1-vinyl-cyclohexanecarboxylic acid

A solution of the commercially available4-oxo-1-vinyl-cyclohexanecarboxylic acid ethyl ester (provided by SmallMolecules Inc.), (1.20 g, 6.11 mmol) in 1,4-dioxane (20 mL) was treatedwith a solution of lithium hydroxide monohydrate (734 mg, 30.6 mmol) inwater (10 mL). After stirring at 50° C. for 2 h, the volatiles wereremoved in vacuo. The residue was partitioned between water and diethylether. The aqueous layer was acidified to pH 1 by addition of saturatedpotassium hydrogen sulfate solution and was extracted with ethyl acetate(3×). The organic layer was dried over anhydrous magnesium sulfate,filtered and the volatiles were removed in vacuo to afford the titlecompound (1.00 g, 97%) as a clear gum. ¹H NMR (300 MHz, CDCl₃)δ1.92-2.08 (m, 2H), 2.36-2.59 (m, 6H), 5.34 (d, J=17.5 Hz, 1H), 5.35 (d,J=10.5 Hz, 1H), 5.97 (dd, J=17.5, 10.5 Hz, 1H), 11.30 (br s, 1H).

Compound 42b.1-((E)-2-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-oxo-cyclohexanecarboxylicacid

A solution of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-methyl-carbamic acidtert-butyl ester (200 mg, 0.55 mmol),4-oxo-1-vinyl-cyclohexanecarboxylic acid (138 mg, 0.82 mmol),palladium(II) acetate (25 mg, 0.11 mmol), tri-(o-tolyl)phosphine (50 mg,0.16 mmol) in anhydrous 1,4-dioxane (2 mL) was treated withtriethylamine (267 μL, 1.91 mmol). After stirring at 100° C. undermicrowave irradiation for 30 minutes, the reaction was cooled to roomtemperature and the volatiles were removed in vacuo. The residue waspartitioned between ethyl acetate and a 5% citric acid solution, theaqueous layer was extracted with ethyl acetate (2 x), the organics werecombined and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 25 g Biotage cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to1:1 to afford the title compound (235 mg, 94%) as a white solid. ¹H NMR(300 MHz, CDCl₃) δ1.48 (s, 9H), 1.68 (d, J=7.1 Hz, 3H), 2.07-2.22 (m,2H), 2.44-2.70 (m, 6H), 2.75 (br s, 3H), 5.61 (br s, 1H), 6.68 (ABq,Δδ_(AB)=0.31, J_(AB)=16.3 Hz, 2H), 7.36 (d, J=8.5 Hz, 1H), 7.60 (d,J=8.5 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 8.05-8.12 (m, 2H). LCMS (m/z)453.1 [M+H], Tr=2.48 min.

Compound 42c.1-((E)-2-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-cis-hydroxy-cyclohexanecarboxylicacid

A cooled (−50° C.) solution of1-((E)-2-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-oxo-cyclohexanecarboxylicacid (235 mg, 0.52 mmol) in anhydrous tetrahydrofuran (15 mL) wastreated with potassium tri-sec-butylborohydride (1.56 mL, 1.56 mmol, 1 Min tetrahydrofuran). The temperature was raised to −30° C. Afterstirring for 20 minutes, the reaction was quenched by addition of asaturated ammonium chloride solution (10 mL). The temperature was raisedto room temperature, the pH adjusted to pH 2 by addition of citric acidthen extracted with ethyl acetate (3×). The organics were combined andthe volatiles were removed in vacuo. The residue was purified by silicagel chromatography using a 25 g Biotage cartridge eluted with acontinuous gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to affordthe title compound (185 mg, 78%) as a white solid. ¹H NMR (300 MHz,CDCl₃) δ1.48 (s, 9H), 1.52-1.62 (m, 4H), 1.67 (d, J=6.9 Hz, 3H),1.95-2.05 (m, 2H), 2.42-2.55 (m, 2H), 2.73 (br s, 3H), 3.73 (br s, 1H),5.57 (br s, 1H), 6.58 (ABq, Δδ_(AB)=0.29, J_(AB)=16.3 Hz, 2H), 7.34 (d,J=8.5 Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 7.72 (d, J=8.5 Hz, 1H), 8.01-8.10(m, 2H). LCMS (m/z) 455.1 [M+H], Tr=2.31 min.

Compound 42d.1-((E)-2-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-cis-(tert-butyl-dimethyl-silanyloxy)-cyclohexanecarboxylicacid

A solution of1-((E)-2-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-cis-hydroxy-cyclohexanecarboxylicacid (185 mg, 0.41 mmol) and imidazole (138 mg, 2.03 mmol) in anhydrousin N,N-dimethylformamide (5 mL) was treated with tert-butyldimethylsilylchloride (153 mg, 1.02 mmol). After stirring for 16 h, the reaction wasquenched by addition of a potassium carbonate solution (10 mL). Afterstirring for 3 h, the pH was adjusted to pH 3 by addition of potassiumhydrogen sulfate solution then extracted with ethyl acetate (3×). Theorganics were combined and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 25 g Biotagecartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate1:0 to 7:3 to afford the title compound (142 mg, 61%) as a white solid.¹H NMR (300 MHz, CDCl₃) δ0.08 (s, 6H), 0.91 (s, 9H), 1.49 (s, 9H),1.53-1.64 (m, 4H), 1.67 (d, J=6.9 Hz, 3H), 1.80-1.92 (m, 2H), 2.38-2.51(m, 2H), 2.72 (br s, 3H), 3.70 (br s, 1H), 5.59 (br s, 1H), 6.58 (ABq,Δδ_(AB)=0.30, J_(AB)=16.3 Hz, 2H), 7.33 (d, J=8.5 Hz, 1H), 7.58 (dd,J=8.5, 1.3 Hz, 1H), 7.71 (d, J=8.5 Hz, 1H), 8.00-8.06 (m, 2H). LCMS(m/z) 569.2 [M+H], Tr=4.37 min.

Compound 42e.(S)-1-((S)-2-{(S)-2-[1-((E)-2-{7-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-2-yl}-vinyl)-4-cis-(tert-butyl-dimethyl-silanyloxy)-cyclohexanecarbonyloxy]-3-methyl-butyrylamino}-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of1-((E)-2-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-cis-(tert-butyl-dimethyl-silanyloxy)-cyclohexanecarboxylicacid (142 mg, 0.25 mmol),(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (108 mg, 0.25 mmol), triethylamine(0.087 mL, 0.62 mmol) and 4-dimethylaminopyridine (15 mg, 0.12 mmol) inanhydrous dichloromethane (10 mL) was treated with2-methyl-6-nitrobenzoic anhydride (146 mg, 0.42 mmol). After stirringfor 60 h, the reaction was quenched by addition of a sodium bicarbonatesolution (10 mL), the aqueous layer was extracted with dichloromethane(2×), the organics were combined and the volatiles were removed invacuo. The residue was purified by silica gel chromatography using a 25g Biotage cartridge eluted with a continuous gradient ofiso-hexanes/ethyl acetate 1:0 to 1:1 to afford the title compound (105mg, 43%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ0.10 (s, 6H), 0.93(s, 9H), 1.01 (d, J=6.9 Hz, 3H), 1.02 (d, J=6.9 Hz, 3H), 1.27 (d, J=6.9Hz, 3H), 1.40-1.50 (m, 9H), 1.68 (d, J=6.9 Hz, 3H), 1.72-1.81 (m, 4H),1.82-1.94 (m, 4H), 1.95-2.10 (m, 2H), 2.25-2.32 (m, 2H), 2.37-2.44 (m,1H), 2.46-2.57 (m, 2H), 2.80 (br s, 3H), 3.52-3.70 (m, 1H), 3.74-3.85(m, 2H), 4.73-4.80 (m, 1H), 5.01 (d, J=4.7 Hz, 1H), 5.30-5.45 (m, 2H),6.70 (ABq, Δδ_(AB)=0.28, J_(AB)=16.3 Hz, 2H), 7.37 (d, J=8.5 Hz, 1H),7.73-7.85 (m, 2H), 7.97 (s, 1H), 8.22 (d, J=8.5 Hz, 1H).

Compound 42

A solution of(S)-1-((S)-2-{(S)-2-[1-((E)-2-{7-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-2-yl}-vinyl)-4-cis-(tert-butyl-dimethyl-silanyloxy)-cyclohexanecarbonyloxy]-3-methyl-butyrylamino}-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (105 mg, 0.11 mmol) in tetrahydrofuran(10 mL) was treated with a solution of potassium carbonate (221 mg, 1.60mmol) in water (5 mL). After stirring at room temperature for 3 h, thepH was adjusted to pH 3 by addition of dilute hydrochloric acid thenextracted with ethyl acetate (3×). The organics were combined and thevolatiles were removed in vacuo. The residue was dissolved indichloromethane (10 mL) and treated with hydrochloric acid in1,4-dioxane (4 M, 5 mL). After stirring at room temperature for 1 h, thevolatiles were removed in vacuo. The residual solvent was azeotroped offwith toluene (3×15 mL) to give crude(S)-1-{(S)-2-[(S)-2-(4-cis-hydroxy-1-{(E)-2-[7-((R)-1-methylamino-ethyl)-quinolin-2-yl]-vinyl}-cyclohexanecarbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid. This residue was dissolved in N,N-dimethylformamide (4 mL) andN,N-diisopropylethylamine (0.093 mL, 0.53 mmol) and added via syringepump over 1 h to a stirred solution of2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (61 mg, 0.16 mmol) in dichloromethane(50 mL) at room temperature under nitrogen. The reaction was quenched byaddition of a sodium bicarbonate solution (10 mL), the aqueous layer wasextracted with dichloromethane (2×), The organics were combined and thevolatiles were removed in vacuo. The residue was purified by preparativereverse phase HPLC to afford the title compound (13 mg, 19%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ1.02 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.7Hz, 3H), 1.27-1.48 (m, 3H), 1.54 (d, J=7.1 Hz, 3H), 1.72 (d, J=7.6 Hz,3H), 1.78-2.05 (m, 5H), 2.11-2.25 (m, 1H), 2.28-2.40 (m, 1H), 2.50-2.62(m, 1H), 2.71-2.84 (m, 1H), 3.27 (s, 3H), 3.52-3.80 (m, 3H), 4.11-4.24(m, 1H), 4.38-4.51 (m, 1H), 5.16 (d, J=9.1 Hz, 1H), 5.86-5.98 (m, 2H),6.36 (ABq, Δδ_(AB)=0.17, J_(AB)=16.5 Hz, 2H), 7.44 (d, J=8.7 Hz, 1H),7.64-7.72 (m, 2H), 7.78 (d, J=8.5 Hz, 1H), 8.18 (d, J=8.7 Hz, 1H). LCMS(m/z) 620.5 [M+H], Tr=2.09 min.

Examples 43 and 44 Compounds 43 and 44

Compound 43a. [1,4]Dioxane-2-carboxylic acid methyl ester

A solution of [1,4]dioxane-2-carboxylic acid (supplied by Enamine Ltd.),(2.90 g, 22.0 mmol) in N,N-dimethylformamide (15 mL) was stirred at roomtemperature. Potassium carbonate (6.07 g, 44.0 mmol) was added and thesuspension was stirred at 5° C. Iodomethane (6.25 g, 2.75 mL, 44.0 mmol)was added dropwise and the reaction mixture was stirred at roomtemperature for 24 h. Water and brine were added and the mixture wasextracted with diethyl ether. The aqueous layer was saturated withsodium chloride and the mixture was extracted with ethyl acetate (3×).The organic extracts were combined, dried over anhydrous magnesiumsulfate, filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 4:1 to 1:1 toafford the title compound (3.06 g, 95%) as a volatile, pale yellow oil.¹H NMR (300 MHz, CDCl₃) δ3.68-3.78 (m, 4H), 3.81 (s, 3H), 3.97-4.05 (m,2H), 4.29 (dd, J=8.5, 3.1 Hz, 1H). LCMS (m/z) 169.2 [M+Na], Tr=0.69 min.

Compound 43b. 2-(1-Hydroxy-ethyl)-[1,4]dioxane-2-carboxylic acid methylester

A solution of N,N-diisopropylamine (1.67 g, 2.3 mL, 16.5 mmol) inanhydrous tetrahydrofuran (10 mL) was stirred at −78° C. under nitrogen.n-Butyllithium (6.6 mL, 16.5 mmol, 2.5 M solution in hexane) was addeddropwise and the reaction mixture was stirred at −78° C. for 30 minutes.A solution of [1,4]dioxane-2-carboxylic acid methyl ester (1.61 g, 11mmol) in tetrahydrofuran (10 mL) was added and the reaction mixture wasstirred at −78° C. for 30 minutes. Acetaldehyde (1.45 g, 1.8 mL, 33mmol) was added in one portion. The cooling bath was removed and thereaction mixture was stirred at room temperature for 30 minutes. Thereaction mixture was cooled to 5° C. and ice-cold 1 M hydrochloric acidwas added to acidify the reaction mixture to pH 2. Sodium chloride wasadded to saturate the solution and the mixture was extracted withdiethyl ether (4×). The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 1:1 to 0:1 toafford the title compound (1.49 g, 71%), as an oil, and as a 3:1 mixtureof diastereoisomers. LCMS (m/z) 213.2 [M+H], Tr=0.62 min.

Compound 43c.2-[1-(Toluene-4-sulfonyloxy)-ethyl]-[1,4]dioxane-2-carboxylic acidmethyl ester

A solution of 2-(1-hydroxy-ethyl)-[1,4]dioxane-2-carboxylic acid methylester (1.49 g, 7.8 mmol) in pyridine (8 mL) was stirred at roomtemperature. 4-Toluenesulfonyl chloride (1.50 g, 7.8 mmol) was added andthe reaction mixture was stirred at room temperature for 18 h. Themajority of the pyridine was evaporated, 1 M hydrochloric acid was addedto acidify the mixture to pH 2 and the mixture was extracted with ethylacetate. The organic extracts were combined and washed with brine. Theorganic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 3:1 to 0:1 toafford the title compound (1.70 g, 63%) as a yellow gum and as a 3:1mixture of diastereoisomers. LCMS (m/z) 367.0 [M+Na], Tr=2.44 min.

Compound 43d. 2-Vinyl-[1,4]dioxane-2-carboxylic acid methyl ester

A solution of2-[1-(toluene-4-sulfonyloxy)-ethyl]-[1,4]dioxane-2-carboxylic acidmethyl ester (1.68 g, 4.9 mmol) in 1,8-diazabicycloundec-7-ene (3 mL)was heated in a sealed tube at 130° C. for 2 h. The reaction mixture wascooled to room temperature and 2 M hydrochloric acid and diethyl etherwere added. The mixture was extracted with diethyl ether and the organicextracts were combined and washed with brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofpentane/diethyl ether 3:1 to 1:1 to afford the title compound (308 mg,36%) as a volatile, colorless oil. ¹H NMR (300 MHz, CDCl₃) δ3.47 (d,J=11.6 Hz, 1H), 3.62-3.78 (m, 2H), 3.81-3.86 (m, 1H), 3.84 (s, 3H),3.99-4.07 (m, 1H), 4.33 (d, J=11.6 Hz, 1H), 5.34 (dd, J=10.7, 1.1 Hz,1H), 5.52 (dd, J=17.4, 1.1 Hz, 1H), 5.79 (dd, J=17.4, 10.7 Hz, 1H). LCMS(m/z) 195.1 [M+Na], Tr=1.23 min.

Compound 43e.2-{(E)-2-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}[1,4]dioxane-2-carboxylicacid methyl ester

A solution of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester (316 mg, 0.9 mmol), 2-vinyl-[1,4]dioxane-2-carboxylicacid methyl ester (154 mg, 0.9 mmol), palladium(II) acetate (41 mg, 0.18mmol), tri(o-tolyl)phosphine (54 mg, 0.18 mmol) andN,N-dicyclohexylmethylamine (351 mg, 0.38 mL, 1.8 mmol) in acetonitrile(8 mL) was heated at 120° C. in a microwave reactor for 30 minutes. Thereaction mixture was filtered and the solvent was evaporated. Theresidue was dissolved in dichloromethane; brine and ice-cold 1 Mhydrochloric acid were added to acidify the mixture to pH 2. The organiclayer was separated and washed with brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 7:3 to 6:4 to afford the title compound (157mg, 40%) as a yellow gum and as a 1:1 mixture of diastereoisomers. LCMS(m/z) 443.1 [M+H], Tr=2.34 min.

Compound 43f.2-{(E)-2-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carboxylicacid

A solution of2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carboxylicacid methyl ester (157 mg, 0.35 mmol) in tetrahydrofuran (6 mL) wasstirred at 5° C. under nitrogen. A solution of lithium hydroxidemonohydrate (30 mg, 0.71 mmol) in water (1.5 mL) was added and thereaction mixture was stirred at room temperature for 18 h. The majorityof the organic solvent was evaporated. The solution was acidified to pH2 with 2 M hydrochloric acid and the mixture was extracted withdichloromethane. The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated to afford the title compound (149 mg, 100%)as a pale yellow foam and as a 1:1 mixture of diastereoisomers. LCMS(m/z) 429.1 [M+H], Tr=1.87 min.

Compound 43g.(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carboxylicacid (50 mg, 0.1 mmol),[(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (44 mg, 0.1 mmol),N,N-diisopropylethylamine (32 mg, 0.043 mL, 0.25 mmol),4-dimethylaminopyridine (6 mg, 0.05 mmol) and 2-methyl-6-nitrobenzoicanhydride (58 mg, 0.17 mmol) in dichloromethane (5 mL) was stirred atroom temperature under nitrogen for 18 h. The reaction mixture wasdiluted with dichloromethane and the solution was washed with ice-coldsaturated ammonium chloride solution, ice-cold saturated sodium hydrogencarbonate solution and brine. The organic solution was filtered througha hydrophobic frit and the filtrate was evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 1:1 to 1:3 to afford the title compound (47mg, 55%) as a white solid and as a 1:1 mixture of diastereoisomers. LCMS(m/z) 842.2/844.0 [M+H], Tr=3.25 and 3.31 min.

Compounds 43 and 44

A solution of(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (69 mg, 0.081 mmol) in tetrahydrofuran(4 mL) was stirred at 0° C. under nitrogen. An ice-cold aqueous solutionof sodium hydroxide (0.1 M, 0.82 mL, 0.082 mmol) was added and thereaction mixture was stirred at 0° C. for 20 minutes. Cold 1 Mhydrochloric acid was added to acidify the mixture to pH 2 and thesolvent was evaporated. The residue was co-evaporated withtetrahydrofuran/toluene (1:1, 3×) and the residue was triturated withdiethyl ether (2×) and the resulting solid was dried to afford(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (0.081 mmol) as a yellow solid and as a 1:1 mixture ofdiastereoisomers which was used crude in the next reaction. LCMS (m/z)712.3 [M+H], Tr=2.46 min.

A mixture of crude(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (0.081 mmol) in 4 M hydrogen chloride in 1,4-dioxane (2 mL) wasstirred at room temperature for 1 h. The solvent was evaporated and theresidue was co-evaporated with diethyl ether (2×) and the resultingsolid was dried to afford(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.081 mmol) as an off-white solid and as a 1:1mixture of diastereoisomers which was used crude in the next reaction.LCMS (m/z) 612.1 [M+H], Tr=1.38 min.

A suspension of crude(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-vinyl}-[1,4]dioxane-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.081 mmol) in dichloromethane (75 mL) was stirredat 0° C. under nitrogen. A solution of N,N-diisopropylethylamine (42 mg,0.324 mmol) in dichloromethane (6 mL) was added and the resultingsolution was stirred at 0° C.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (62 mg, 0.162 mmol) was added and thereaction mixture was stirred at 0° C. for 30 minutes and then at roomtemperature for 1 h. The majority of the solvent was evaporated and thesolution (˜20 mL) was washed with ice-cold 1 M hydrochloric acid andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:4 to 0:1to afford a 1:1 mixture of diastereoisomers. The mixture was purified byreverse phase preparative HPLC eluting with acetonitrile/water 3:7 toafford the two separate diastereoisomers.

Compound 43 (First eluting) Diastereoisomer 1: (4.0 mg, 8%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ 1.06 (d, J=6.7 Hz, 3H), 1.12 (d, J=6.7Hz, 3H), 1.56-1.60 (m, 7H), 1.68-1.73 (m, 1H), 1.95-2.00 (m, 1H),2.20-2.35 (m, 2H), 2.69-2.78 (m, 1H), 3.55-3.85 (m, 5H), 4.03-4.12 (m,1H), 4.43-4.48 (m, 1H), 4.57 (d, J=11.6 Hz, 1H), 5.12 (q, J=6.7 Hz, 1H),5.24 (d, J=10.4 Hz, 1H), 5.39 (d, J=8.5 Hz, 1H), 5.83 (q, J=7.2 Hz, 1H),6.29 (d, J=16.3 Hz, 1H), 6.97 (d, J=16.3 Hz, 1H), 7.49 (d, J=8.5 Hz,1H), 7.61 (dd, J=8.5, 1.3 Hz, 1H), 7.86-7.90 (m, 2H), 8.28 (d, J=8.5 Hz,1H). LCMS (m/z) 594.2 [M+H], Tr=2.14 min.

Compound 44 (Second eluting) Diastereoisomer 2: (4.6 mg, 10%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ1.08 (d, J=6.5 Hz, 3H), 1.10 (d, J=6.5Hz, 3H), 1.49-1.54 (m, 1H), 1.59 (d, J=6.7 Hz, 3H), 1.64 (d, J=7.1 Hz,3H), 1.66-1.71 (m, 1H), 1.90-1.96 (m, 1H), 2.20-2.28 (m, 2H), 2.63-2.71(m, 1H), 3.55-3.93 (m, 6H), 4.40-4.47 (m, 2H), 4.98 (d, J=12.0 Hz, 1H),5.09 (q, J=6.7 Hz, 1H), 5.20 (d, J=8.9 Hz, 1H), 5.79 (q, J=7.1 Hz, 1H),6.10 (d, J=16.3 Hz, 1H), 6.88 (d, J=16.3 Hz, 1H), 7.47 (d, J=8.5 Hz,1H), 7.62 (dd, J=8.5, 1.6 Hz, 1H), 7.85-7.88 (m, 2H), 8.26 (d, J=8.5 Hz,1H). LCMS (m/z) 594.2 [M+H], Tr=2.17 min.

Examples 45 and 46 Compounds 45 and 46

Compound 45a. Morpholine-2,4-dicarboxylic acid 4-tert-butyl ester2-methyl ester

A solution of morpholine-2,4-dicarboxylic acid 4-tert-butyl ester(supplied by NeoMPS, Inc.), (5.78 g, 25.0 mmol) in N,N-dimethylformamide(30 mL) was subsequently treated with potassium carbonate (4.14 g, 30.0mmol) and iodomethane (7.10 g, 3.1 mL, 50.0 mmol). After stirring atroom temperature for 18 h, the reaction mixture was diluted with waterand the aqueous layer was extracted with ethyl acetate. The organicextracts were combined, washed with water (4×) and brine then filteredthrough a hydrophobic frit and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 3:1 to 1:3 to afford the title compound (4.83g, 79%) as a white solid. ¹H NMR (300 MHz, CDCl₃) δ1.49 (s, 9H),3.06-3.15 (m, 2H), 3.56-3.64 (m, 1H), 3.75-3.79 (m, 1H), 3.81 (s, 3H),4.00-4.14 (m, 3H). LCMS (m/z) 268.1 [M+Na], Tr=1.94 min.

Compound 45b. 2-(1-Hydroxy-ethyl)-morpholine-2,4-dicarboxylic acid4-tert-butyl ester 2-methyl ester

A cooled (−78° C.) solution of N,N-diisopropylamine (1.3 mL, 9.092 mmol,dried over calcium hydride) in anhydrous tetrahydrofuran (10 mL) wastreated with a solution of n-butyllithium in hexanes (3.4 mL, 8.524mmol, 2.5 M). After stirring at −78° C. for 30 minutes, the mixture wastreated with a solution of morpholine-2,4-dicarboxylic acid 4-tert-butylester 2-methyl ester (1.394 g, 5.683 mmol) in anhydrous tetrahydrofuran(10 mL). After stirring at −78° C. for 20 minutes, the mixture wastreated with acetaldehyde (0.96 mL, 17.05 mmol). After stirring at roomtemperature for 1 h, the reaction was quenched at 0° C. withhydrochloric acid (1 M, 40 mL). The aqueous layer was extracted withdichloromethane (2×). The organics were combined, filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to3:2 to afford the title compound (1.476 g, 90%) as a yellow oil and as a2:1 mixture of diastereoisomers.

Compound 45c.2-(1-Trifluoromethanesulfonyloxy-ethyl)-morpholine-2,4-dicarboxylic acid4-tert-butyl ester 2-methyl ester

A cooled (−78° C.) suspension of sodium hydride (262 mg, 6.542 mmol, 60%in mineral oil) in anhydrous tetrahydrofuran (10 mL) was treated with asolution of 2-(1-hydroxy-ethyl)-morpholine-2,4-dicarboxylic acid4-tert-butyl ester 2-methyl ester (1.262 g, 4.361 mmol) in anhydroustetrahydrofuran (10 mL). The light yellow suspension was stirred at −78°C. for 15 minutes and then treated with a solution ofN-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (3.425 g, 8.722mmol) in anhydrous tetrahydrofuran (10 mL). After stirring at roomtemperature for 1.5 h, the reaction was quenched with a saturatedsolution of ammonium chloride. The aqueous layer was extracted withdichloromethane (2×). The organics were combined, filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to4:1 to afford the title compound (1.384 g, 75%) as a light yellow oiland as a 2:1 mixture of diastereoisomers.

Compound 45d. 2-Vinyl-morpholine-2,4-dicarboxylic acid 4-tert-butylester 2-methyl ester

A warm (35° C.) solution of2-(1-trifluoromethanesulfonyloxy-ethyl)-morpholine-2,4-dicarboxylic acid4-tert-butyl ester 2-methyl ester (1.384 g, 3.284 mmol) indichloromethane (40 mL) was treated with1,8-diazabicyclo[5.4.0]undec-7-ene (2.5 mL, 16.422 mmol). After stirringat 35° C. for 2 h, the reaction was cooled to 0° C. and quenched withhydrochloric acid (1 M, 50 mL). The aqueous layer was extracted withdichloromethane. The organics were combined, filtered through a phaseseparator and the volatiles were removed in vacuo to provide the titlecompound as an orange oil which was used without further purification inthe next step. ¹H NMR (300 MHz, CDCl₃) δ1.49 (s, 9H), 3.13-3.29 (m, 2H),3.60-3.73 (m, 1H), 3.79 (s, 3H), 3.83-3.92 (m, 2H), 4.24 (d, J=13.1 Hz,1H), 5.37 (d, J=10.9 Hz, 1H), 5.55 (d, J=17.4 Hz, 1H), 5.84 (dd, J=17.4,10.9 Hz, 1H).

Compound 45e. 2-Vinyl-morpholine-2,4-dicarboxylic acid 4-tert-butylester

A solution of crude 2-vinyl-morpholine-2,4-dicarboxylic acid4-tert-butyl ester 2-methyl ester (3.284 mmol) intetrahydrofuran/methanol/water (50 mL, 2:2:1) was treated with lithiumhydroxide monohydrate (413.3 mg, 9.852 mmol). After stirring at roomtemperature for 2 h, the volatiles were removed in vacuo and the residuewas cooled to 0° C. and quenched with hydrochloric acid (1 M). Theaqueous layer was extracted with dichloromethane (2×). The combinedorganics were filtered through a phase separator and the volatiles wereremoved in vacuo to provide the title compound (761.4 mg, 90% over 2steps) as colorless needles. ¹H NMR (300 MHz, CDCl₃) δ1.48 (s, 9H),3.31-3.42 (m, 1H), 3.45-3.61 (m, 2H), 3.87-3.94 (m, 2H), 4.02 (d, J=13.6Hz, 1H), 5.44 (d, J=10.7 Hz, 1H), 5.60 (d, J=17.4 Hz, 1H), 5.85 (dd,J=17.4, 10.7 Hz, 1H).

Compound 45f.2-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-morpholine-2,4-dicarboxylicacid 4-tert-butyl ester

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (95 mg, 0.178 mmol),2-vinyl-morpholine-2,4-dicarboxylic acid 4-tert-butyl ester (45.7 mg,0.178 mmol), tri(o-tolyl)phosphine (11 mg, 0.036 mmol) and triethylamine(0.08 mL, 0.534 mmol) in 1,4-dioxane (5 mL) was degassed by bubblingnitrogen through for 5 minutes then warmed to 50° C. and treated withtris(dibenzylideneacetone)dipalladium(0) (16.3 mg, 0.018 mmol). Afterstirring at 100° C. for 50 minutes the reaction was cooled to roomtemperature, filtered through a pad of Celite, which was rinsed withethyl acetate. The volatiles were removed in vacuo to provide crudetitle compound as an orange foam that was used without furtherpurification. LCMS (m/z) 711.3 [M+H], Tr=2.14 min.

Compound 45 and 46

A solution of crude2-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-morpholine-2,4-dicarboxylicacid 4-tert-butyl ester (0.178 mmol), 4-dimethylaminopyridine (21.7 mg,0.178 mmol) and triethylamine (0.08 mL, 0.534 mmol) in drydichloromethane (150 mL) was treated with 2-methyl-6-nitrobenzoicanhydride (122.6 mg, 0.356 mmol). After stirring at room temperature for1.5 h, more 2-methyl-6-nitrobenzoic anhydride (122.6 mg, 0.356 mmol) wasadded. After stirring at room temperature for 16 h, more triethylamine(0.2 mL) and 4-dimethylaminopyridine (217 mg, 1.780 mmol) were added.After stirring at room temperature for 4 h, the volatiles were removedin vacuo. The residue was partitioned between dichloromethane and asaturated solution of sodium bicarbonate. The aqueous layer wasextracted with dichloromethane. The organics were combined, filteredthrough a phase separator and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 25 g Isolutecartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate1:0 to 0:1 then by reverse phase preparative HPLC eluting with acontinuous gradient of water/acetonitrile 95:5 to 0:1 to afford thetitle compound as two separate isomers.

Compound 45 (Second eluting) Diastereomer 1 (1.9 mg, 1.5%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ1.06 (d, J=6.7 Hz, 6H), 1.47-1.54 (m,10H), 1.55-1.77 (m, 8H), 1.90-2.00 (m, 1H), 2.16-2.31 (m, 2H), 2.62-2.74(m, 1H), 3.04-3.18 (m, 2H), 3.54-3.62 (m, 1H), 3.68-3.80 (m, 1H),3.83-3.93 (m, 1H), 3.94-4.03 (m, 1H), 4.38-4.47 (m, 1H), 4.57-4.67 (m,1H), 5.03 (d, J=12.0 Hz, 1H), 5.10 (q, J=6.7 Hz, 1H), 5.19 (d, J=8.7 Hz,1H), 5.80 (q, J=7.3 Hz, 1H), 6.18 (d, J=16.3 Hz, 1H), 6.93 (d, J=16.3Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.62 (dd, J=8.5, 1.3 Hz, 1H), 7.87 (d,J=8.5 Hz, 1H), 7.90 (s, 1H), 8.27 (d, J=8.5 Hz, 1H). LCMS (m/z) 693.3[M+H], Tr=2.68 min.

Compound 46 (First eluting) Diastereomer 2 (0.8 mg, 1%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ0.97-1.06 (m, 6H), 1.40 (d, J=6.7 Hz,3H), 1.43-1.58 (m, 13H), 1.65-1.74 (m, 1H), 1.97-2.07 (m, 1H), 2.18-2.37(m, 2H), 3.42-3.49 (m, 1H), 3.74-4.03 (m, 4H), 5.16 (q, J=7.1 Hz, 1H),5.27-5.33 (m, 1H), 5.35 (q, J=6.7 Hz, 1H), 6.58 (d, J=16.3 Hz, 1H), 6.78(d, J=7.1 Hz, 1H), 7.20 (d, J=16.0 Hz, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.70(d, J=8.5 Hz, 1H), 7.74-7.81 (m, 1H), 7.99 (s, 1H), 8.06 (d, J=7.6 Hz,1H), 8.13 (d, J=8.5 Hz, 1H). LCMS (m/z) 693.5 [M+H], Tr=3.39 min.

Example 47 Compound 47

Compound 47a.(S)-1-{(S)-2-[(S)-2-((E)-4-{2-[(R)-1-(tert-Butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of(E)-4-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid methyl ester (193 mg, 0.47 mmol) in 1,4-dioxane (10 mL) was treatedwith a solution of lithium hydroxide monohydrate (112 mg, 4.7 mmol) inwater (5 mL). After stirring at 50° C. for 1 h, the volatiles wereremoved in vacuo. The residue was partitioned between water and diethylether. The aqueous layer was acidified to pH 2 by addition ofhydrochloric acid then extracted with ethyl acetate (3×). The organicswere combined and the volatiles were removed in vacuo. The residue wasdissolved in dichloromethane (10 mL) and was treated with4-dimethylaminopyridine (57 mg, 0.47 mmol), 2-methyl-6-nitrobenzoicanhydride (275 mg, 0.80 mmol), and triethylamine (0.197 mL, 1.41 mmol).After stirring for 30 min(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (203 mg, 0.47 mmol) was added. Afterstirring for 16 h, the reaction was quenched by addition of a sodiumbicarbonate solution (10 mL), the aqueous layer was extracted withdichloromethane (2×), the organics were combined and the volatiles wereremoved in vacuo. The residue was purified by silica gel chromatographyusing a 25 g Biotage cartridge eluted with a continuous gradient ofiso-hexanes/acetone 1:0 to 3:1 to afford the title compound (260 mg,68%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ0.97-1.05 (m, 6H), 1.29(d, J=6.9 Hz, 3H), 1.44 (br s, 9H), 1.55 (s, 3H), 1.57 (s, 3H), 1.68 (d,J=6.9 Hz, 3H), 1.71-2.00 (m, 4H), 2.02-2.30 (m, 3H), 2.80 (br s, 3H),3.56 (br s, 1H), 3.78-3.83 (m, 1H), 4.75-4.82 (m, 1H), 4.93-5.01 (m,2H), 5.35-5.45 (m, 1H), 6.65-6.82 (m, 2H), 7.36 (d, J=8.5 Hz, 1H),7.72-7.83 (m, 2H), 7.98 (s, 1H), 8.21 (d, J=8.5 Hz, 1H). LCMS (m/z)814.1 [M+H], Tr=3.97 min.

Compound 47

A solution of(S)-1-{(S)-2-[(S)-2-((E)-4-{2-[(R)-1-(tert-butoxycarbonyl-methyl-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (260 mg, 0.32 mmol) in tetrahydrofuran(30 mL) was prepared and a solution of potassium carbonate (663 mg, 4.80mmol) in water (15 mL) was added. The reaction was stirred at roomtemperature for 1.5 h. The reaction mixture was acidified to pH 3 withhydrochloric acid (1 M) then extracted with ethyl acetate (3×20 mL). Thecombined extracts were dried over anhydrous sodium sulfate, filtered andevaporated. The residue was dried under vacuum for 1 h before dissolvingin dichloromethane (30 mL) and treating with hydrogen chloride in1,4-dioxane (4 M, 15 mL). It was stirred for 1 h at room temperaturethen evaporated. Residual water was azeotroped off with toluene (2×20mL) to yield a white solid (251 mg). The white solid was dissolved inanhydrous N,N-dimethylformamide (10 mL) and N,N-diisopropylethylamine(279 μL, 1.6 mmol) was added. This solution was then added via a syringepump to a stirred solution of2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (183 mg, 0.48 mmol) in anhydrousdichloromethane (150 mL) over 1 h. The reaction mixture was washed withsaturated sodium bicarbonate solution (30 mL), filtered through a phaseseparator and evaporated. The residue was purified by reverse phasepreparative HPLC to yield the title product (54 mg, 30%) as a whitesolid. ¹H NMR (300 MHz, CDCl₃) δ0.97 (s, 3H), 0.99 (s, 3H), 1.44 (s,3H), 1.50-1.55 (m, 6H), 1.56-2.03 (m, 4H), 1.68 (d, J=7.4 Hz, 3H),2.20-2.34 (m, 1H), 2.64-2.79 (m, 1H), 3.14 (s, 3H), 3.90-4.01 (m, 1H),4.50-4.61 (m, 1H), 4.85 (d, J=8.5 Hz, 3H), 5.88-6.15 (m, 1H), 6.30 (d,J=16.5 Hz, 1H), 6.38 (d, J=8.0 Hz, 1H), 6.53 (d, J=16.3 Hz, 1H),6.61-6.87 (m, 1H), 7.28 (d, J=8.5 Hz, 1H), 7.52 (d, J=8.3 Hz, 1H), 7.68(d, J=8.5 Hz, 1H), 7.87 (s, 1H), 8.05 (d, J=8.5 Hz, 1H). LCMS (m/z)564.2 [M+H], Tr=2.54 min.

Example 48 Compound 48

Compound 48a. Ethyl 2,2-bis(hydroxymethyl)-3-oxobutanoate

To a stirred solution of ethyl acetoacetate (150 g, 1.2 mol) informaldehyde (210 mL, 2.8 mol, 40% aq.) and dioxane (550 mL) was addeddropwise triethylamine (1.0 M in tetrahydrofuran, 57 mL, 0.06 mol) at 0°C. The temperature was raised up to 35-40° C. and the mixture wasstirred for 1 h. The reaction mixture was diluted with water (400 mL),and the side products were extracted with toluene (3×600 mL). Theaqueous phase was evaporated at 35° C. to one fourth of the initialvolume and extracted with ethyl acetate (3×900 mL). The extracts weredried over sodium sulfate and evaporated to get the crude product (75 g)as yellow oil, which was used directly to the next step withoutpurification.

Compound 48b. Ethyl 5-acetyl-1,3-dioxane-5-carboxylate

A solution of ethyl 2,2-bis(hydroxymethyl)-3-oxobutanoate (75 g, 0.39mol) and 40% formaldehyde (360 mL) in concentrated hydrochloric acid(360 mL) was heated at 50° C. for 17 h. The two-phase solution wascooled to room temperature and extracted with ethyl acetate (2×200 mL).The combined organic phases were washed with water, dried over magnesiumsulfate, and evaporated to give a residue, which was purified by flashcolumn chromatography eluting with a continuous gradient of petroleumether/ethyl acetate 15:1 to 9:1 to afford the title compound (33 g, 14%,2 steps) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ1.25 (m, 3H), 2.31(s, 3H), 4.25-4.19 (m, 4H), 4.35 (d, J=12.0 Hz, 2H), 4.77 (d, J=6.0 Hz,1H), 4.82 (d, J=6.0 Hz, 1H).

Compound 48c. Ethyl 5-(1-hydroxyethyl)-1,3-dioxane-5-carboxylate

Ethyl 5-acetyl-1,3-dioxane-5-carboxylate (2) (22.3 g, 0.11 mol) wasdissolved in ethanol (100 mL) and sodium borohydride (2.08 g, 0.055 mol)was added at 0° C. in portions. The mixture was stirred at the sametemperature for 1 h. The excess of sodium borohydride was destroyed withacetone and the solution was extracted with ethyl acetate (3×600 mL).The combined organic extracts were concentrated in vacuo to give thetitle compound (18.9 g, 84%) as colorless oil.

Compound 48d. Ethyl 5-(1-(tosyloxy)ethyl)-1,3-dioxane-5-carboxylate

To a mixture of ethyl 5-(1-hydroxyethyl)-1,3-dioxane-5-carboxylate (16.8g, 83 mmol) in dry pyridine (40 mL) at 0° C. was added 4-toluenesulfonylchloride (19.1 g, 99 mmol). The mixture was stirred at rt for 16 h.Water (40 mL) was added, and the mixture was extracted with ethylacetate (2×400 mL). The organic layer was washed with brine (3×). Afterconcentrated in vacuo, the crude product was purified by silica gelchromatography eluting with petroleum ether/ethyl acetate 25:1 to 5:1 toprovide the title compound (22.7 g, 76%) as a colorless oil. ¹H NMR (400MHz, CDCl₃) δ1.26 (m, 3H), 1.34 (d, J=6.8 Hz, 3H), 2.45 (s, 3H), 3.80(d, J=11.6 Hz, 1H), 3.87 (d, J=11.6 Hz, 1H), 4.02 (d, J=11.6 Hz, 1H),4.08 (d, J=12.0 Hz, 1H), 4.20-4.17 (m, 2H), 5.03-4.98 (m, 1H), 7.34 (d,J=8.0 Hz, 2H), 7.79 (d, J=8.8 Hz, 2H).

Compound 48e. 5-Vinyl-[1,3]dioxane-5-carboxylic acid ethyl ester

A mixture of ethyl 5-(1-(tosyloxy)ethyl)-1,3-dioxane-5-carboxylate (20.7g, 58 mmol) and DBU (25 mL) was heated at 140° C. for 3 h. After beingcooled to rt, the reaction mixture was poured into ether (300 mL), andthe organic layer was washed with brine (3×). The ether layer was driedover sodium sulfate, and concentrated in vacuo. The residue was purifiedby silica gel chromatography eluting with petroleum ether/ether 30:1 to20:1 to give a crude product, which was distilled under reduced pressure(45-49° C./0.3 mbar) to afford the title compound (3.6 g, 33%) ascolorless oil. ¹H NMR (400 MHz, CDCl₃) δ1.29 (t, J=11.0 Hz, 3H), 3.70(d, J=11.6 Hz, 2H), 4.25 (t, J=7.2 Hz, 2H), 4.44 (t, J=11.2 Hz, 2H),4.71 (d, J=8.0 Hz, 1H), 4.95 (d, J=6.0 Hz, 1H), 5.24 (m, 1H), 5.27 (m,1H), 5.65 (m, 1H). This ¹H spectral data is in good agreement with thatreported for the synthesis of the title compound, the which is describedin Borremans, F. et al. Bull. Soc. Chim. Belg. 1976, 85, 681-696.

Compound 48f. 5-Vinyl-[1,3]dioxane-5-carboxylic acid

A solution of 5-vinyl-[1,3]dioxane-5-carboxylic acid ethyl ester (397.7mg, 2.136 mmol) in tetrahydrofuran/methanol/water (15 mL, 2:2:1) wastreated with lithium hydroxide monohydrate (269 mg, 6.408 mmol). Afterstirring at room temperature for 1.5 h, the volatiles were removed invacuo and the residue was cooled to 0° C. and quenched with hydrochloricacid (1 M). The aqueous layer was saturated with sodium chloride,extracted with dichloromethane (3×). The combined organics were filteredthrough a phase separator and the volatiles were removed in vacuo toprovide the title compound (320.3 mg, 95%) as a colourless solid. ¹H NMR(300 MHz, CDCl₃) δ3.72 (d, J=11.6 Hz, 2H), 4.50 (d, J=11.4 Hz, 2H), 4.73(d, J=6.2 Hz, 1H), 5.02 (d, J=6.0 Hz, 1H), 5.27-5.39 (m, 2H), 5.69 (dd,J=17.6, 10.9 Hz, 1H).

Compound 48g.5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,3]dioxane-5-carboxylicacid

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (398.4 mg, 0.745 mmol),5-vinyl-[1,3]dioxane-5-carboxylic acid (117.9 mg, 0.745 mmol),tri(o-tolyl)phosphine (45.3 mg, 0.149 mmol) and triethylamine (0.32 mL,2.235 mmol) in 1,4-dioxane (15 mL) was degassed by bubbling nitrogenthrough for 5 minutes then warmed to 50° C. and treated withtris(dibenzylideneacetone)dipalladium(0) (68.2 mg, 0.074 mmol). Afterstirring at 100° C. for 40 minutes the reaction was cooled to roomtemperature, filtered through a pad of Celite, which was rinsed withethyl acetate. The volatiles were removed in vacuo to provide crudetitle compound as an orange foam that was used without furtherpurification. LCMS (m/z) 612.2 [M+H], Tr=1.70 min.

Compound 48

A solution of crude5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,3]dioxane-5-carboxylicacid (0.745 mmol) in dry dichloromethane (18 mL) was added via syringepump to a solution of 2-methyl-6-nitrobenzoic anhydride (385.0 mg, 1.117mmol), 4-dimethylaminopyridine (273.0 mg, 2.235 mmol) in drydichloromethane (230 mL) containing 4 Å molecular sieves over 3 h. Afterthe end of the addition, the reaction was stirred at room temperaturefor 40 minutes then filtered and the volatiles were partially removed invacuo. The organics were washed with pH 4 citrate buffer, a saturatedsolution of sodium bicarbonate and filtered on a phase separator. Thevolatiles were removed in vacuo and the residue was purified by silicagel chromatography using a 25 g Isolute cartridge eluted with acontinuous gradient of iso-hexanes/acetone 1:0 to 1:1 to provide thetitle compound at 90% purity (133.8 mg, 30% yield). A third of thismaterial was purified by reverse phase preparative HPLC eluting with acontinuous gradient of water/acetonitrile 95:5 to 0:1 to afford thetitle compound (8.1 mg) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.06(d, J=6.7 Hz, 3H), 1.11 (d, J=6.7 Hz, 3H), 1.47-1.80 (m, 8H), 2.12-2.31(m, 2H), 2.65-2.77 (m, 1H), 3.59-3.66 (m, 1H), 3.85 (d, J=11.4 Hz, 1H),3.92 (d, J=11.4 Hz, 1H), 4.39-4.53 (m, 2H), 4.63-4.71 (m, 1H), 4.98 (d,J=6.2 Hz, 1H), 5.08 (q, J=6.9 Hz, 1H), 5.35 (d, J=9.1 Hz, 1H), 5.82 (q,J=7.3 Hz, 1H), 6.15 (d, J=16.5 Hz, 1H), 6.63 (d, J=16.5 Hz, 1H), 7.47(d, J=8.5 Hz, 1H), 7.69 (dd, J=8.5, 1.8 Hz, 1H), 7.75 (s, 1H), 7.86 (d,J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 594.1 [M+H], Tr=2.60min.

Example 49 Compound 49

Compound 49a. 2-(1-Hydroxy-ethyl)-tetrahydro-pyran-2-carboxylic acidmethyl ester

A solution of N,N-diisopropylamine (909 mg, 1.25 mL, 9 mmol) intetrahydrofuran (4 mL) was stirred at −78° C. under nitrogen.n-Butyllithium (3.6 mL, 9 mmol, 2.5 M solution in hexane) was addeddropwise and the reaction mixture was stirred at −78° C. for 30 minutes.A solution of tetrahydro-pyran-2-carboxylic acid methyl ester (864 mg, 6mmol) in tetrahydrofuran (8 mL) was added and the reaction mixture wasstirred at −78° C. for 20 minutes. Acetaldehyde (792 mg, 1.0 mL, 18mmol) was added in one portion. The cooling bath was removed and thereaction mixture was stirred at room temperature for 1 h. The reactionmixture was cooled to 0° C. and ice-cold hydrochloric acid (2 M) wasadded to acidify the reaction mixture to pH 2. Sodium chloride was addedto saturate the aqueous phase and the mixture was extracted with diethylether. The organic extracts were combined and washed with brine. Theorganic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 1:1 to 0:1 toafford the title compound (1.28 g) as an oil and as a 2:1 mixture ofdiastereoisomers. LCMS (m/z) 189.3 [M+H], Tr=1.23 min.

Compound 49b. 2-Vinyl-tetrahydro-pyran-2-carboxylic acid methyl ester

A suspension of sodium hydride (200 mg, 5 mmol, 60% dispersion in oil)in tetrahydrofuran (6 mL) was stirred at −78° C. under nitrogen. Asolution of 2-(1-hydroxy-ethyl)-tetrahydro-pyran-2-carboxylic acidmethyl ester (620 mg, 3.3 mmol) in tetrahydrofuran 3 mL) was added andthe reaction mixture was stirred at −78° C. for 15 minutes. A solutionof N-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (2.60 g, 6.6mmol) in tetrahydrofuran (10 mL) was added and the reaction mixture wasstirred at −78° C. for 30 minutes. The reaction mixture was warmed toroom temperature and then stirred at room temperature for 1 h. Thereaction mixture was cooled to 5° C. and saturated ammonium chloridesolution was cautiously added. The reaction mixture was extracted withdiethyl ether. The organics were combined and washed with brine. Theorganic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 10:1 to 5:1 toafford2-(1-trifluoromethanesulfonyloxy-ethyl)-tetrahydro-pyran-2-carboxylicacid methyl ester (2.12 g, 3.3 mmol) as an oil and as a 2:1 mixture ofdiastereoisomers which was used immediately in the next step. LCMS (m/z)321.0 [M+H], Tr=2.88 min.

A solution of2-(1-trifluoromethanesulfonyloxy-ethyl)-tetrahydro-pyran-2-carboxylicacid methyl ester (2.12 g, 3.3 mmol) in dichloromethane (5 mL) wasstirred at room temperature under nitrogen. 1,8-Diazabicycloundec-7-ene(2.0 g, 2 mL, 13.2 mmol) was added and the reaction mixture was stirredat room temperature for 18 h. The reaction mixture was cooled to 0° C.and acidified to pH 2 with ice cold hydrochloric acid (2 M). The mixturewas extracted with dichloromethane. The organic extracts were combinedand washed with brine. The organic solution was filtered through ahydrophobic frit and the filtrate was evaporated. The residue waspurified by silica gel chromatography using a gradient ofpentane/diethyl ether 10:1 to 5:1 to afford the title compound (466 mg,75% over two steps) as an oil. ¹H NMR (300 MHz, CDCl₃) δ 1.55-1.80 (m,5H), 2.21-2.29 (m, 1H), 2.68-2.77 (m, 1H), 3.79 (s, 3H), 3.91-3.98 (m,1H), 5.26 (dd, J=10.7, 0.9 Hz, 1H), 5.41 (dd, J=17.4, 1.1 Hz, 1H), 5.87(dd, J=17.4, 10.7 Hz, 1H). LCMS (m/z) 193.2 [M+H], Tr=2.01 min.

Compound 49c.(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloroethyl ester

A solution of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester (320 mg, 0.9 mmol),2-vinyl-tetrahydro-pyran-2-carboxylic acid methyl ester (330 mg, 0.9mmol), tris(dibenzylideneacetone)dipalladium(0) (82 mg, 0.09 mmol),tri(o-tolyl)phosphine (54 mg, 0.18 mmol) and N,N-dicyclohexylmethylamine(526 mg, 0.58 mL, 2.7 mmol) in acetonitrile (4 mL) was heated at 120° C.in a microwave reactor for 30 minutes. The reaction mixture was filteredand the solvent was evaporated. The residue was dissolved in ethylacetate and the solution was washed with ice-cold hydrochloric acid (1M). The aqueous layer was extracted with ethyl acetate and the organicextracts were combined and washed with brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated toafford2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carboxylicacid methyl ester (130 mg, 21%) as an oil LCMS (m/z) 441.1 [M+H],Tr=2.75 min.

A solution of2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carboxylicacid methyl ester (130 mg, 0.3 mmol) in tetrahydrofuran (6 mL) wasstirred at 5° C. under nitrogen. A solution of lithium hydroxidemonohydrate (25 mg, 0.6 mmol) in water (1.5 mL) was added and thereaction mixture was stirred at room temperature for 18 h. The majorityof the organic solvent was evaporated. The solution was acidified to pH2 with ice-cold hydrochloric acid (2 M) and the mixture was extractedwith dichloromethane. The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated to afford2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carboxylicacid (135 mg, 0.3 mmol) as a yellow gum which was used crude in the nextstep. LCMS (m/z) 427.1 [M+H], Tr=2.17 min.

A solution of2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carboxylicacid (135 mg, 0.3 mmol), N,N-diisopropylethylamine (96 mg, 0.133 mL,0.75 mmol), 4-dimethylaminopyridine (80 mg, 0.66 mmol) and2-methyl-6-nitrobenzoic anhydride (193 mg, 0.56 mmol) in dichloromethane(5 mL) was stirred at 0° C. under nitrogen. A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (142 mg, 0.33 mmol) in dichloromethane(2 mL) was added and the reaction mixture was stirred at roomtemperature for 6 h. The reaction mixture was diluted withdichloromethane and the solution was washed with ice-cold saturatedsodium hydrogen carbonate solution, water, ice-cold hydrochloric acid (1M), water and brine. The organic solution was filtered through ahydrophobic frit and the filtrate was evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 3:1 to 1:3 to afford the title compound (44mg, 17%) as a white solid and as a 1:1 mixture of two diastereoisomers.LCMS (m/z) 840.2, 842.0 [M+H], Tr=3.75 min.

Compound 49

A solution of(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloroethyl ester (42 mg, 0.05 mmol) in tetrahydrofuran (2mL) was stirred at 0° C. under nitrogen. An ice-cold aqueous solution ofsodium hydroxide (0.1 M, 0.5 mL, 0.05 mmol) was added and the reactionmixture was stirred at 0° C. for 30 minutes. Additional sodium hydroxidesolution (0.1 M, 1.0 mL, 0.1 mmol) was added and the reaction mixturewas stirred at 0° C. for 1 h. Additional sodium hydroxide solution (0.1M, 1.0 mL, 0.1 mmol) was added and the reaction mixture was stirred at0° C. for 90 minutes. Cold hydrochloric acid (1 M) was added to acidifythe mixture to pH 2 and the solvent was evaporated. The residue wasco-evaporated with tetrahydrofuran/toluene (1:1, 3×) and the residue wastriturated with diethyl ether (2×) and the resulting solid was dried toafford(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (0.05 mmol) as a yellow solid and as a 1:1 mixture ofdiastereoisomers which was used crude in the next reaction. LCMS (m/z)710.3 [M+H], Tr=2.80 min.

A mixture of crude(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid (0.05 mmol) in hydrochloric acid (4 M in 1,4-dioxane, 2 mL) wasstirred at room temperature for 1 h. The solvent was evaporated and theresidue was co-evaporated with diethyl ether (2×) and the resultingsolid was dried to afford(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.05 mmol) as an off-white solid and as a 1:1mixture of diastereoisomers which was used crude in the next reaction.LCMS (m/z) 610.1 [M+H], Tr=1.60 min.

A suspension of crude(S)-1-{(S)-2-[(S)-2-(2-{(E)-2-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-vinyl}-tetrahydro-pyran-2-carbonyloxy)-3-methyl-butyrylamino]-propionyl}-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.05 mmol) in dichloromethane (45 mL) was stirred at0° C. under nitrogen. A solution of N,N-diisopropylethylamine (26 mg,0.2 mmol) in dichloromethane (5 mL) was added and the resulting solutionwas stirred at 0° C. 2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate methanaminium (38 mg, 0.1 mmol) was addedand the reaction mixture was stirred at 0° C. for 30 minutes and then atroom temperature for 1 h. The reaction mixture was cooled to 0° C. andthe solution was washed with ice-cold hydrochloric acid (1 M) and brine.The organic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was triturated with diethyl etherand the residue was dried. The residue purified by silica gelchromatography using a gradient of iso-hexanes/acetone 2:1. The residuewas triturated with iso-hexanes/diethyl ether 1:1 and the resultingsolid was dried to afford the title compound (6.0 mg, 21%) as a whitesolid and as a 1:1 mixture of diastereoisomers. LCMS (m/z) 592.2 [M+H],Tr=2.51 min and (m/z) 592.2 [M+H], Tr=2.61 min.

Examples 50 and 51 Compounds 50 and 51

Compound 50a. Tetrahydro-pyran-3-carboxylic acid benzyl ester

A cooled (0° C.) solution of tetrahydro-pyran-3-carboxylic acid (2.831g, 21.752 mmol) in dichloromethane (60 mL) and N,N-dimethylformamide (5drops) was treated with oxalyl chloride (2.0 mL, 23.927 mmol). Afterstirring at room temperature for 2 h, benzyl alcohol (2.5 mL, 23.927mmol) was added. After stirring at room temperature for 2 h and standingovernight, the volatiles were removed in vacuo and the residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to4:1 to afford the title compound (4.587 g, 96%) as a white solid. ¹H NMR(300 MHz, CDCl₃) δ 1.58-1.88 (m, 3H), 2.03-2.13 (m, 1H), 2.62-2.72 (m,1H), 3.47 (ddd, J=11.4, 9.6, 3.4 Hz, 1H), 3.63 (dd, J=11.4, 9.1 Hz, 1H),3.85 (app dt, J=11.1, 3.8 Hz, 1H), 4.06 (ddd, J=11.4, 4.0, 1.6 Hz, 1H),5.12, 5.17 (ABq, J_(AB)=12.5 Hz, 2H), 7.31-7.46 (m, 5H).

Compound 50b. 3-(1-Hydroxy-ethyl)-tetrahydro-pyran-3-carboxylic acidbenzyl ester

A cooled (−78° C.) solution of N,N-diisopropylamine (4.7 mL, 33.316mmol, dried over calcium hydride) in anhydrous tetrahydrofuran (60 mL)was treated with a solution of n-butyllithium in hexanes (12.5 mL,31.234 mmol, 2.5 M). After stirring at −78° C. for 25 minutes, themixture was treated with a solution of tetrahydro-pyran-3-carboxylicacid benzyl ester (4.587 g, 20.823 mmol) in anhydrous tetrahydrofuran(20 mL). After stirring at −78° C. for 15 minutes, the mixture wastreated with acetaldehyde (3.5 mL, 62.469 mmol). After stirring at roomtemperature for 45 minutes, the reaction was quenched at 0° C. withhydrochloric acid (2 M, 50 mL). The aqueous layer was saturated withsodium chloride, extracted with dichloromethane (2 x). The organics werecombined, filtered through a phase separator and the volatiles wereremoved in vacuo. The residue was purified by silica gel chromatographyusing a 100 g Isolute cartridge eluted with a continuous gradient ofiso-hexanes/ethyl acetate 1:0 to 3:2 to afford the title compound (2.212g, 40%) as a yellow oil and as a 3:1 mixture of diastereoisomers. LCMS(m/z) 287.0 [M+Na], Tr=2.00 min.

Compound 50c.3-(1-Trifluoromethanesulfonyloxy-ethyl)-tetrahydro-pyran-3-carboxylicacid benzyl ester

A cooled (−78° C.) suspension of sodium hydride (502.1 mg, 12.552 mmol,60% in mineral oil) in anhydrous tetrahydrofuran (20 mL) was slowlytreated with a solution of3-(1-hydroxy-ethyl)-tetrahydro-pyran-3-carboxylic acid benzyl ester(2.212 g, 8.368 mmol) in anhydrous tetrahydrofuran (20 mL). The lightyellow suspension was stirred at −78° C. for 15 minutes and then treatedwith a solution ofN-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (6.572 g, 16.736mmol) in anhydrous tetrahydrofuran (20 mL). After stirring at roomtemperature for 3 h, more sodium hydride (502.1 mg, 12.552 mmol, 60% inmineral oil) was added. After stirring at room temperature for 2 h, moresodium hydride (502.1 mg, 12.552 mmol, 60% in mineral oil) was added.After stirring at room temperature for 30 minutes, the reaction wasquenched at −20° C. with hydrochloric acid (2 M, 50 mL). The aqueouslayer was extracted with dichloromethane (3×). The organics werecombined, filtered through a phase separator and the volatiles wereremoved in vacuo to afford the title compound in a mixture which wasused without further purification. LCMS (m/z) 397.0 [M+H], Tr=2.48 min.

Compound 50d. 3-Vinyl-tetrahydro-pyran-3-carboxylic acid benzyl ester

A solution of3-(1-trifluoromethanesulfonyloxy-ethyl)-tetrahydro-pyran-3-carboxylicacid benzyl ester (8.368 mmol) in dichloromethane (100 mL) was treatedwith 1,8-diazabicyclo[5.4.0]undec-7-ene (5.0 mL, 33.472 mmol). Afterstirring at room temperature for 2.5 h, the volatiles were removed invacuo and the residue was purified by silica gel chromatography using a100 g Isolute cartridge eluted with a continuous gradient ofiso-hexanes/ethyl acetate 1:0 to 4:1 to afford the title compound (809.5mg, 39% over two steps) as a colorless oil. ¹H NMR (300 MHz, CDCl₃)δ1.58-1.80 (m, 3H), 2.29-2.40 (m, 1H), 3.42-3.54 (m, 2H), 3.81 (app dt,J=10.5, 4.2 Hz, 1H), 4.27 (dd, J=11.4, 1.8 Hz, 1H), 5.13-5.27 (m, 4H),5.77 (dd, J=17.4, 10.9 Hz, 1H), 7.31-7.42 (m, 5H).

Compound 50e. 3-Vinyl-tetrahydro-pyran-3-carboxylic acid

A solution of 3-vinyl-tetrahydro-pyran-3-carboxylic acid benzyl ester(809.5 mg, 3.286 mmol) in tetrahydrofuran/methanol/water (25 mL, 2:2:1)was treated with lithium hydroxide monohydrate (413.6 mg, 9.859 mmol).After stirring at room temperature for 2 h, more lithium hydroxidemonohydrate (413.6 mg 9.859 mmol) was added. After stirring at roomtemperature for 2 h, the volatiles were removed in vacuo and the residuewas cooled to 0° C. and quenched with hydrochloric acid (1 M). Theaqueous layer was saturated with sodium chloride, extracted withdichloromethane (3×). The combined organics were filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 25 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to1:1 to afford crude title compound which was dissolved in diethyl ether.Organics were extracted with a saturated solution of sodium bicarbonate.The aqueous layer was acidified with hydrochloric acid (2 M) to pH ˜2,saturated with sodium chloride and extracted with dichloromethane (2×)to provide the title compound (377.1 mg, 73%) as a white solid. ¹H NMR(300 MHz, CDCl₃) δ1.59-1.90 (m, 3H), 2.27-2.38 (m, 1H), 3.39-3.55 (m,2H), 3.87 (app dt, J=11.2, 4.0 Hz, 1H), 4.26 (dd, J=11.4, 2.0 Hz, 1H),5.22-5.32 (m, 2H), 5.79 (dd, J=17.8, 10.7 Hz, 1H).

Compound 50f.3-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-tetrahydro-pyran-3-carboxylicacid

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (600 mg, 1.122 mmol),3-vinyl-tetrahydro-pyran-3-carboxylic acid (175.3 mg, 1.122 mmol),tri(o-tolyl)phosphine (68.3 mg, 0.224 mmol) and triethylamine (0.47 mL,3.366 mmol) in 1,4-dioxane (15 mL) was degassed by bubbling nitrogenthrough for 5 minutes then warmed to 50° C. and treated withtris(dibenzylideneacetone)dipalladium(0) (102.7 mg, 0.112 mmol). Afterstirring at 100° C. for 45 minutes the reaction was cooled to roomtemperature, filtered through a pad of Celite, which was rinsed withethyl acetate. The volatiles were removed in vacuo to provide crudetitle compound as an orange foam that was used without furtherpurification. LCMS (m/z) 610.2 [M+H], Tr=1.97 min.

Compounds 50 and 51

A solution of crude3-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-tetrahydro-pyran-3-carboxylicacid (1.122 mmol) in dry dichloromethane (20 mL) was added via syringepump to a solution of 2-methyl-6-nitrobenzoic anhydride (772.6 mg, 2.244mmol), 4-dimethylaminopyridine (548.2 mg, 4.488 mmol) in drydichloromethane (350 mL) containing 4 Å molecular sieves over 4 h. Afterthe end of the addition, the reaction was stirred at room temperaturefor 3 days then filtered and the volatiles were partially removed invacuo. The organics were washed with pH 4 citrate buffer, a saturatedsolution of sodium bicarbonate and filtered on a phase separator. Thevolatiles were removed in vacuo and the residue was purified by silicagel chromatography using a 50 g Isolute cartridge eluted with acontinuous gradient of iso-hexanes/acetone 1:0 to 2:3 to provide twofractions that were further purified by reverse preparative HPLC elutingwith water/acetonitrile 95:5 to 0:1 to afford two separate, singleisomers 1 and 2.

Compound 50 (First eluting) Diastereomer 1 (50.1 mg, 7.5%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ1.0 (d, J=6.4 Hz, 3H), 1.09 (d, J=6.7Hz, 3H), 1.47-1.72 (m, 9H), 1.73-1.88 (m, 2H), 1.91-2.02 (m, 1H),2.11-2.32 (m, 2H), 2.35-2.45 (m, 1H), 2.65-2.78 (m, 1H), 3.46-3.67 (m,3H), 3.84-3.94 (m, 1H), 4.39-4.54 (m, 2H), 5.03-5.16 (m, 2H), 5.34 (d,J=9.1 Hz, 1H), 5.81 (q, J=7.3 Hz, 1H), 6.23 (d, J=16.5 Hz, 1H), 6.62 (d,J=16.5 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.66 (dd, J=8.5, 1.3 Hz, 1H),7.75 (s, 1H), 7.85 (d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z)592.2 [M+H], Tr=2.31 min.

Compound 51 (Second eluting) Diastereomer 2 (28.8 mg, 4.3%) as a whitesolid. ¹H NMR (300 MHz, CD₃OD) δ1.06 (d, J=6.7 Hz, 3H), 1.09 (d, J=6.7Hz, 3H), 1.60 (d, J=6.7 Hz, 3H), 1.65 (d, J=7.3 Hz, 3H), 1.68-1.79 (m,3H), 1.88-2.00 (m, 2H), 2.12-2.29 (m, 2H), 2.55-2.76 (m, 2H), 3.49-3.67(m, 3H), 3.84-3.93 (m, 1H), 4.29 (dd, J=11.2, 1.6 Hz, 1H), 4.39-4.48 (m,1H), 5.08 (q, J=6.7 Hz, 1H), 5.28 (d, J=8.7 Hz, 1H), 5.82 (q, J=7.1 Hz,1H), 6.20 (d, J=16.7 Hz, 1H), 6.62 (d, J=16.7 Hz, 1H), 7.46 (d, J=8.5Hz, 1H), 7.66 (dd, J=8.5, 1.6 Hz, 1H), 7.79 (s, 1H), 7.85 (d, J=8.5 Hz,1H), 8.25 (d, J=8.5 Hz, 1H). LCMS (m/z) 592.1 [M+H], Tr=2.27 min.

Example 52 Compound 52

Compound 52a. (S)-Cyclobutyl-hydroxy-acetic acid

A cooled (0° C.) solution of cyclobutyl-L-glycine (572.8 mg, 4.435 mmol)in aqueous sulfuric acid (0.5 M, 18 mL) was treated with a solution ofsodium nitrite (1.836 g, 26.610 mmol) in water (10 mL). After stirringat room temperature for 19 h, the solution was saturated with sodiumchloride and extracted with tetrahydrofuran (3×). The organics werefiltered through a hydrophobic frit and the volatiles were removed invacuo. The solid residue was then extracted with ethyl acetate, theorganics were filtered through a hydrophobic frit and the volatiles wereremoved in vacuo to afford the title compound (235.9 mg, 41%) as anorange gum which was used without further purification. ¹H NMR (300 MHz,CDCl₃) δ1.76-2.23 (m, 6H), 2.66-2.82 (m, 1H), 4.18 (d, J=5.8 Hz, 1H).

Compound 52b.(S)-1-[(S)-2-((S)-2-Cyclobutyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

A cooled (0° C.) solution of(S)-2-{(S)-3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester 53a (1.065 g, 1.994 mmol) in dichloromethane (15mL) was treated with a solution of hydrogen chloride (3 mL, 11.964 mmol,4 M in 1,4-dioxane). After stirring at room temperature for 2 h, asolution of hydrogen chloride (3 mL, 11.964 mmol, 4 M in 1,4-dioxane)was added. After stirring for 2 h, the volatiles were removed in vacuo.Residual water was azeotroped off with toluene to provide(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide hydrochloride as a whitesolid which was then combined with (S)-cyclobutyl-hydroxy-acetic acid(235.9 mg, 1.812 mmol) and dry dichloromethane (20 mL). This suspensionwas cooled to 0° C. and subsequently treated withN,N-diisopropylethylamine (0.95 mL, 5.436 mmol) and(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(961.7 mg, 2.174 mmol). The reaction was slowly warmed to roomtemperature. After stirring for 17 h the reaction was quenched at 0° C.with hydrochloric acid (2 M). The aqueous layer was extracted withdichloromethane. The organics were washed with a saturated solution ofsodium bicarbonate. The basic aqueous layer was extracted withdichloromethane. The organics were combined, filtered through a phaseseparator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/acetone 1:0 to 1:1 toafford the title compound (427.5 mg, 39%) as an orange foam. ¹H NMR (300MHz, CDCl₃) δ1.40 (d, J=8.0 Hz, 3H), 1.59 (d, J=6.4 Hz, 3H), 1.63-2.10(m, 10H), 2.21-2.32 (m, 1H), 2.58-2.78 (m, 2H), 3.41-3.54 (m, 1H), 4.01(app t, J=5.8 Hz, 1H), 4.52 (d, J=13.8 Hz, 1H), 5.28 (app pentet, J=6.9Hz, 1H), 5.41 (app pentet, J=8.0 Hz, 1H), 7.00 (d, J=8.2 Hz, 1H), 7.40(d, J=8.5 Hz, 1H), 7.65 (dd, J=8.5, 1.8 Hz, 1H), 7.71 (d, J=8.5 Hz, 1H),8.06 (d, J=7.1 Hz, 1H), 8.16 (d, J=8.5 Hz, 1H), 8.44 (s, 1H). LCMS (m/z)547.0, 549.0 [M+H], Tr=2.39 min.

Compound 52c.(E)-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Cyclobutyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid

A solution of(S)-1-[(S)-2-((S)-2-cyclobutyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (427.5 mg, 0.782 mmol),2,2-dimethyl-but-3-enoic acid (89.3 mg, 0.782 mmol),tri(o-tolyl)phosphine (47.6 mg, 0.156 mmol) and triethylamine (0.33 mL,2.346 mmol) in 1,4-dioxane (10 mL) was degassed by bubbling nitrogenthrough for 5 minutes then warmed to 50° C. and treated withtris(dibenzylideneacetone)dipalladium(0) (71.6 mg, 0.078 mmol). Afterstirring at 100° C. for 1.5 h, tris(dibenzylideneacetone) dipalladium(0)(71.6 mg, 0.078 mmol) and tri(o-tolyl)phosphine (47.6 mg, 0.156 mmol)were added. After stirring for 40 minutes at 100° C., the reaction wascooled to room temperature, filtered through a pad of Celite, which wasrinsed with ethyl acetate. The volatiles were removed in vacuo toprovide crude title compound as an orange foam that was used withoutfurther purification. LCMS (m/z) 580.2 [M+H], Tr=2.01 min.

Compound 52

A solution of crude(E)-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-cyclobutyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid (0.782 mmol) in dry dichloromethane (20 mL) was added via syringepump to a solution of 2-methyl-6-nitrobenzoic anhydride (538.4 mg, 1.564mmol), 4-dimethylaminopyridine (382.1 mg, 3.128 mmol) in drydichloromethane (280 mL) containing 4 Å molecular sieves over 4 h. Afterthe end of the addition, the reaction was stirred at room temperaturefor 30 minutes then filtered and the volatiles were partially removed invacuo. The organics were washed with pH 4 citrate buffer, a saturatedsolution of sodium bicarbonate and filtered on a phase separator. Thevolatiles were removed in vacuo and the residue was purified by silicagel chromatography using a 50 g Isolute cartridge eluted with acontinuous gradient of iso-hexanes/acetone 1:0 to 1:1 and then byreverse phase preparative HPLC eluting with a continuous gradient ofwater/acetonitrile 95:5 to 0:1 to afford the title compound (48.6 mg,11%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.43 (s, 3H), 1.47-1.75(m, 12H), 1.89-2.12 (m, 6H), 2.24-2.33 (m, 1H), 2.69 (app dt, J=12.9,3.1 Hz, 1H), 2.84 (app sextet, J=8.0 Hz, 1H), 3.60 (dd, J=11.6, 2.9 Hz,1H), 4.38-4.47 (m, 1H), 5.10 (q, J=6.7 Hz, 1H), 5.53 (d, J=8.0 Hz, 1H),5.81 (q, J=7.1 Hz, 1H), 6.38 (d, J=16.3 Hz, 1H), 6.57 (d, J=16.3 Hz,1H), 7.44 (d, J=8.5 Hz, 1H), 7.61 (dd, J=8.5, 1.6 Hz, 1H), 7.81-7.87 (m,2H), 8.25 (d, J=8.5 Hz, 1H). LCMS (m/z) 562.1 [M+H], Tr=2.68 min.

Example 53 Compound 53

Compound 53a.((S)-2-{(S)-3-[(R)-1-(7-Bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

A cooled (0° C.) solution of(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (3.007 g, 6.948 mmol) intetrahydrofuran/water (60 mL, 5:1) was treated with lithium hydroxidemonohydrate (874.4 mg, 20.844 mmol). After stirring at 0° C. for 40minutes the reaction was quenched with hydrochloric acid (1 M, 50 mL).The aqueous layer was extracted with ethyl acetate (2×). The organicswere combined, dried over anhydrous sodium sulfate, filtered and thevolatiles were removed in vacuo. Residual trichlorethanol was azeotropedoff with toluene (3×) to provide(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-carboxylicacid as a white solid which was then combined with(R)-1-(7-bromo-quinolin-2-yl)-ethylamine hydrochloride (1.998 g, 6.948mmol) and suspended in anhydrous acetonitrile (60 mL) andtetrahydrofuran (10 mL). The suspension was cooled to 0° C. andsubsequently treated with N,N-diisopropylethylamine (6 mL, 34.740 mmol)and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (3.699 g, 9.727 mmol). After slowly warmingto room temperature and stirring for 16 h, the reaction was quenched at0° C. with hydrochloric acid (1 M, 70 mL). The aqueous layer wasextracted with ethyl acetate (2×). The organics were combined, washedwith a saturated solution of sodium bicarbonate, dried over anhydroussodium sulfate, filtered and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 100 g Isolutecartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate1:0 to 1:4 to afford the title compound (3.702 g, 99%) as a white foam.¹H NMR (300 MHz, CDCl₃) δ1.38 (d, J=6.9 Hz, 3H), 1.43 (s, 9H), 1.47-1.55(m, 2H), 1.58 (d, J=6.9 Hz, 3H), 1.60-1.78 (m, 2H), 2.22-2.31 (m, 1H),2.65-2.78 (m, 1H), 3.39-3.52 (m, 1H), 4.55 (d, J=13.4 Hz, 1H), 5.18-5.34(m, 2H), 5.36-5.45 (m, 1H), 7.37 (d, J=8.5 Hz, 1H), 7.65 (dd, J=8.5, 1.6Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 8.04 (d, J=6.5 Hz, 1H), 8.14 (d, J=8.5Hz, 1H), 8.36 (s, 1H). LCMS (m/z) 536.1, 537.1 [M+H], Tr=2.58 min.

Compound 53b.(S)-1-[(S)-2-(2-Cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

((S)-2-{(S)-3-[(R)-1-(7-Bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester, (1.07 g, 2.00 mmol) was suspended indichloromethane (20 mL) and the mixture was cooled to 0° C. withstirring. 4 M Hydrogen chloride in 1,4-dioxane (11 mL) was added and thereaction mixture stirred for 90 minutes and then evaporated. Residualwater was azeotroped off with toluene and the residue was suspended indichloromethane (20 mL) and cyclopropyl-hydroxy-acetic acid (255 mg,2.20 mmol) was added. The mixture was cooled to 0° C. with stirring andN,N-diisopropylethylamine (775 mg, 1.05 mL, 6.00 mmol) was addedfollowed by (benzotriazol-1-yloxy)tris(dimethylamino) phosphoniumhexafluorophosphate (1.06 g, 2.40 mmol). The reaction was allowed towarm to ambient temperature and stirred for 18 h. 1M Hydrochloric acidwas added and the organic layer was washed with saturated sodiumbicarbonate solution, passed through a hydrophobic frit and evaporated.The residue was purified by silica gel chromatography eluting with agradient of ethyl acetate/methanol 1:0 to 9:1 to afford the titlecompound (328 mg) as a white foam. The original aqueous extracts werere-extracted with dichloromethane/methanol (9:1, 2×) and the organicextracts dried over anhydrous sodium sulfate, filtered and evaporated.The residue purified by silica gel chromatography eluting with agradient of ethyl acetate/methanol 1:0 to 17:3 to afford the titlecompound (378 mg) as a white foam. The two foams were combined (706 mg,66%). ¹H NMR (300 MHz, CDCl₃) δ0.46-0.73 (m, 4H), 1.04-1.18 (m, 1H),1.49-1.59 (m, 6H), 1.53-1.78 (m, 2H), 1.95-2.04 (m, 1H), 2.25-2.34 (m,1H), 2.64-2.82 (m, 1H), 3.44-3.55 (m, 2H), 3.60-3.84 (m, 1H), 4.50-4.60(m, 1H), 5.23-5.33 (m, 1H), 5.48-5.55 (m, 1H), 7.36-7.41 (m, 1H),7.62-7.73 (m, 2H), 8.13-8.19 (m, 2H), 8.47-8.51 (m, 1H). LCMS (m/z)[M+H] 532.0, 534.0 Tr=2.07 min. LCMS (m/z) 532.0, 533.9 [M+H], Tr=2.12min.

Compound 53

A solution of(S)-1-[(S)-2-(2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (592 mg, 1.11 mmol),5-vinyl[1,3]dioxane-5-carboxylic acid (176 mg, 1.11 mmol), triethylamine(343 mg, 472 μL, 3.39 mmol) and tri(o-tolyl)phosphine (67 mg, 0.22 mmol)in 1,4-dioxane (15 mL) was degassed with nitrogen for 5 minutes and thenwarmed to 50° C. under nitrogen with stirring.Tris(dibenzylideneacetone)dipalladium(0) (101 mg, 0.11 mmol) was addedand the mixture was heated to reflux for 20 minutes and then allowed tocool to room temperature. The suspension was filtered through Celite andthe filtrate evaporated to give crude5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-(2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,3]dioxane-5-carboxylicacid.

To a stirred mixture of powdered 4 Å molecular sieves,2-methyl-6-nitrobenzoic anhydride (764 mg, 2.22 mmol) and4-(dimethylamino)-pyridine (542 mg, 4.44 mmol) in dichloromethane (375mL) was added a solution of crude5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-(2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,3]dioxane-5-carboxylicacid in dichloromethane (20 mL) over 5 h via syringe pump. The flaskoriginally containing the crude acid was washed with furtherdichloromethane (5 mL) and this was added to the reaction mixture overca. 5 minutes. The reaction mixture was stirred for 16 h and thenfiltered through Celite and evaporated to ˜100 mL, washed successivelywith ice-cold saturated ammonium chloride solution, ice-cold saturatedsodium bicarbonate solution, saturated brine and then filtered through ahydrophobic frit. The residue was purified by silica gel chromatographyeluting with a gradient of iso-hexanes/acetone 1:0 to 1:1 and then byreverse phase preparative HPLC using a gradient of acetonitrile/water1:9 to 3:2 modified with 0.1% formic acid to give the title compound (39mg, 6%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD) δ0.52-0.76 (m, 4H),1.30-1.40 (m, 1H), 1.49-1.76 (m, 2H), 1.60 (d, J=6.9 Hz, 3H), 1.64 (d,J=7.1 Hz, 3H), 1.93-2.02 (m, 1H), 2.21-2.30 (m, 1H), 2.66-2.77 (m, 1H),3.59-3.66 (m, 1H), 3.85, 3.96 (ABq, J_(AB)=11.4 Hz, 2H), 4.41-4.51 (m,2H), 4.63-4.70 (m, 1H), 4.82 (d, J=6.2 Hz, 1H), 4.98 (d, J=6.0 Hz, 1H),5.04-5.13 (m, 2H), 5.83-5.91 (m, 1H), 6.17 (d, J=16.7 Hz, 1H), 6.66 (d,J=16.7 Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.68 (dd, J=8.5, 1.5 Hz, 1H),7.83 (s, 1H), 7.86 (d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z)592.2 [M+H], Tr=2.04 min.

Example 54 Compound 54

Compound 54a. (2S,3S)-2-Hydroxy-3-methoxy-butyric acid

A solution of methyl L-allo-threonine (900 mg, 6.75 mmol) in 1 Msulphuric acid (10 mL) was prepared and cooled to 0° C., before adding asolution of sodium nitrite (693 mg, 10 mmol) in water (2.25 mL) dropwiseover 10 minutes. The reaction mixture was stirred at 0° C. for 15 h.Solid ammonium chloride was added to saturate the solution, which wasthen extracted with diethyl ether (3×25 mL). The extracts were combinedand dried over anhydrous sodium sulfate, filtered and evaporated to givethe title product (193 mg, 21%) as a colorless oil. ¹H NMR (300 MHz,CDCl₃) δ1.29 (d, J=6.5 Hz, 3H), 3.48 (s, 3H), 3.71-3.81 (m, 1H), 4.35(d, J=4.7 Hz, 1H).

Compound 54b.(S)-1-[(S)-2-((2S,3S)-2-Hydroxy-3-methoxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

A solution of((S)-2-{(S)-3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester (838 mg, 1.44 mmol) in dichloromethane (20 mL) wastreated with hydrogen chloride in 1,4-dioxane (4 M, 1.44 mL, 5.76 mmol)and stirred for 2 h at room temperature. The solution was evaporated todryness. The residue was dissolved in dichloromethane (20 mL) and(2S,3S)-2-hydroxy-3-methoxy-butyric acid (193 mg, 1.44 mmol) was addedfollowed by N,N-diisopropylethylamine (1.0 mL, 5.76 mmol) andO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(819 mg, 2.16 mmol). The solution was stirred at room temperature for 20h and then evaporated to dryness. The residue was purified by silica gelchromatography eluting with a gradient of acetone/ethyl acetate 1:9 to1:1 to yield the title compound (243 mg, 31%) as a white gum. ¹H NMR(300 MHz, CDCl₃) δ1.14 (d, J=6.2 Hz, 3H), 1.33-1.50 (m, 6H), 1.58 (d,J=6.7 Hz, 3H), 2.20-2.30 (m, 1H), 2.66-2.79 (m, 1H), 3.26-3.63 (m, 3H),3.37 (s, 3H), 3.90 (d, J=11.8 Hz, 1H), 4.20 (d, J=5.1 Hz, 1H), 4.46-4.56(m, 1H), 5.20-5.31 (m, 1H), 5.36-5.49 (m, 1H), 7.41 (d, J=8.5 Hz, 1H),7.61-7.70 (m, 2H), 8.15 (d, J=8.5 Hz, 1H), 8.41 (s, 1H). LCMS (m/z)550.0, 552.0 [M+H], Tr=2.05 min.

Compound 54c.(E)-4-{2-[(R)-1-({(S)-1-[(S)-2-((2S,3S)-2-Hydroxy-3-methoxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid

A solution of(S)-1-[(S)-2-((2S,3S)-2-hydroxy-3-methoxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (243 mg, 0.44 mmol) and2,2-dimethylbut-3-enoic acid (50 mg, 0.44 mmol) in 1,4-dioxane wasprepared and triethylamine (183 μL, 1.32 mmol) was added. The reactionmixture was purged with nitrogen for 5 minutes thentris(dibenzylideneacetone)dipalladium(0) (40 mg, 0.044 mmol) was added.The reaction mixture was heated at 100° C. under a nitrogen atmospherefor 1.5 h. The reaction was filtered through hyflo supercel, washingthrough with ethyl acetate. The filtrate was evaporated in vacuo toyield the crude title compound as a yellow gum. LCMS (m/z) 584.2 [M+H],Tr=1.77 min.

Compound 54

A solution of 2-methyl-6-nitrobenzoic anhydride (389 mg, 1.13 mmol) inanhydrous dichloromethane (160 mL) was prepared and 4 Å molecular sievesand 4-(dimethylamino)pyridine (275 mg, 2.25 mmol) were added. A solutionof crude(E)-4-{2-[(R)-1-({(S)-1-[(S)-2-((2S,3S)-2-hydroxy-3-methoxy-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2,2-dimethyl-but-3-enoicacid (ca. 0.44 mmol) in dichloromethane (11 mL) was added over 5.5 h viasyringe pump. After the end of addition, the reaction mixture wasstirred for a further 30 minutes at room temperature. The solution wasfiltered and washed with saturated aqueous sodium bicarbonate solution(70 mL). The organics were filtered through a phase separating cartridgeand evaporated to give a yellow gum (0.97 g) which was purified bysilica gel chromatography eluting with a gradient of ethylacetate/acetone 1:0 to 4:1 to yield a colorless gum (133 mg) which waspurified by reverse phase preparative HPLC eluting acetonitrile/water9:11 to yield the title product (15.5 mg, 6%) as a white solid. ¹H NMR(300 MHz, CD₃OD) δ1.23 (d, J=6.3 Hz, 3H), 1.44 (s, 3H), 1.53 (s, 3H),1.58 (d, J=6.9 Hz, 3H), 1.61 (d, J=7.4 Hz, 3H), 1.34-1.84 (m, 2H),1.91-2.01 (m, 2H), 2.25-2.36 (m, 1H), 2.63-2.77 (m, 1H), 3.43 (s, 3H),2.54-3.67 (m, 1H), 2.73-3.83 (m, 1H), 4.40-4.50 (m, 1H), 4.07-5.16 (m,1H), 5.56 (d, J=5.6 Hz, 1H), 5.86 (q, J=7.1 Hz, 1H), 6.45, 6.60 (ABq,J_(AB)=16.1 Hz, 2H), 7.44 (d, J=8.5 Hz, 1H), 7.59 (m, 1H), 7.84 (d,J=8.3 Hz, 1H), 7.93 (s, 1H), 8.25 (d, J=8.5 Hz, 1H). LCMS (m/z) 566.1[M+H], Tr=2.32 min.

Example 55 Compound 55

Compound 55a. (R)-2-Hydroxymethyl-2-methyl-but-3-enoic acid methyl ester

A solution of (R)-2-methoxymethyl-2-methyl-but-3-enoic acid (600 mg, 4.2mmol) in dichloromethane (40 mL) was stirred at 0° C. under nitrogen.Boron tribromide (1 M in dichloromethane, 16.8 mL, 16.8 mmol) was addedand the reaction mixture was stirred at 0° C. for 90 minutes. Methanol(20 mL) was cautiously added and the reaction mixture was warmed to roomtemperature over 30 minutes and then stirred at room temperature for 30minutes. The solvent was evaporated and the residue was purified bysilica gel chromatography using a gradient of pentane/diethyl ether 3:1to 1:1 to afford the title compound (540 mg, 90%) as a clear oil. ¹H NMR(300 MHz, CDCl₃) δ1.35 (s, 3H), 2.33 (m, 1H), 3.62 (m, 1H), 3.75 (s,3H), 3.75-3.80 (m, 1H), 5.21 (dd, J=17.4, 0.5 Hz, 1H), 5.24 (dd, J=10.7,0.5 Hz, 1H), 5.98 (dd, J=17.4, 10.7 Hz, 1H). LCMS (m/z) 145.1 [M+H],Tr=1.16 min.

Compound 55b. (R)-2-Difluoromethoxymethyl-2-methyl-but-3-enoic acidmethyl ester

A solution of (R)-2-hydroxymethyl-2-methyl-but-3-enoic acid methyl ester(288 mg, 2.0 mmol) in acetonitrile (8 mL) was stirred at roomtemperature under nitrogen. Copper(I) iodide (80 mg, 0.4 mmol) was addedfollowed by difluoro-fluorosulfonyl-acetic acid (356 mg, 0.2 mL, 2.0mmol) and the reaction mixture was heated at reflux for 30 minutes. Thereaction mixture was cooled to room temperature, water was added and themixture was extracted with ethyl acetate. The organic extracts werecombined and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography using a gradient ofpentane/diethyl ether 10:1 to 5:1 to afford the title compound (195 mg,50%) as a clear oil. ¹H NMR (300 MHz, CDCl₃) δ1.40 (s, 3H), 3.75 (s,3H), 3.86 (d, J=9.4 Hz, 1H), 4.10 (d, J=9.4 Hz, 1H), 5.24 (d, J=17.4 Hz,1H), 5.24 (dd, J=10.9 Hz, 1H), 5.95 (dd, J=17.4, 10.9 Hz, 1H), 6.22 (t,J=74.3 Hz, 1H).

Compound 55c. (R)-2-Difluoromethoxymethyl-2-methyl-but-3-enoic acid

A solution of (R)-2-difluoromethoxymethyl-2-methyl-but-3-enoic acidmethyl ester (159 mg, 0.8 mmol) in tetrahydrofuran (6 mL) was stirred at5° C. under nitrogen. A solution of lithium hydroxide monohydrate (100mg, 2.4 mmol) in water (1.5 mL) was added and the reaction mixture wasstirred at 5° C. for 1 h and then at room temperature for 20 h. Waterwas added and the mixture was acidified to pH 2 with hydrochloric acid(2 M) and the mixture was extracted with diethyl ether. The organicextracts were combined and washed with brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated toafford the title compound (133 mg, 92%) as a clear oil. ¹H NMR (300 MHz,d₆-DMSO) 61.24 (s, 3H), 3.80 (d, J=9.4 Hz, 1H), 4.02 (d, J=9.4 Hz, 1H),5.19 (dd, J=10.6 Hz, 1H), 5.20 (d, J=17.6 Hz, 1H), 5.91 (dd, J=17.6,10.6 Hz, 1H), 6.68 (t, J=75.6 Hz, 1H), 12.5-13.0 (br s, 1H). LCMS (m/z)179.1 [M−H], Tr=1.79 min.

Compound 55

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (160 mg, 0.3 mmol),(R)-2-difluoromethoxymethyl-2-methyl-but-3-enoic acid (54 mg, 0.3 mmol),tri(o-tolyl)phosphine (18 mg, 0.06 mmol) and triethylamine (91 mg, 0.125mL, 0.9 mmol) in 1,4-dioxane (6 mL) was stirred at 50° C. undernitrogen. Tris(dibenzylideneacetone)dipalladium(0) (27 mg, 0.03 mmol)was added and the reaction mixture was heated at reflux for 40 minutes.The reaction mixture was cooled to room temperature and the mixture wasfiltered through a hydrophobic frit and the filtrate was evaporated toafford crude(E)-(R)-2-difluoromethoxymethyl-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2-methyl-but-3-enoicacid (0.3 mmol) which was used in the next step. LCMS (m/z) 634.2 [M+H],Tr=2.04 min. A mixture of powdered 4 Å molecular sieves (200 mg),4-dimethylaminopyridine (146 mg, 1.2 mmol), and 2-methyl-6-nitrobenzoicanhydride (206 mg, 0.6 mmol) in dichloromethane (200 mL) was stirred atroom temperature under nitrogen. A solution of crude(E)-(R)-2-difluoromethoxymethyl-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2-methyl-but-3-enoicacid (0.3 mmol) in dichloromethane (4 mL) was added via syringe pumpover 4 h and the reaction mixture was stirred at room temperature for 18h. The reaction mixture was filtered through Celite and the filtrate wasevaporated to a volume of approximately 50 mL. The solution was washedwith ice-cold saturated ammonium chloride solution, ice-cold saturatedsodium hydrogen carbonate solution and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 1:2 0:1 and then by reverse phase preparativeHPLC eluting with a gradient of acetonitrile/water 1:4 to 1:0 to affordthe title compound (21 mg, 11%) as a white solid. ¹H NMR (300 MHz,CD₃OD) δ1.01 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.7 Hz, 3H), 1.60 (d, J=6.7Hz, 3H), 1.61 (s, 3H), 1.66 (d, J=7.1 Hz, 3H), 1.66-1.70 (m, 2H),1.93-1.98 (m, 1H), 2.13-2.26 (m, 2H), 2.64-2.73 (m, 1H), 3.55-3.67 (m,1H), 3.94 (d, J=9.6 Hz, 1H), 4.26 (d, J=9.6 Hz, 1H), 4.40-4.45 (m, 1H),5.04 (d, J=12.0 Hz, 1H), 5.08 (q, J=6.7 Hz, 1H), 5.23 (d, J=9.2 Hz, 1H),5.81 (q, J=7.1 Hz, 1H), 6.24 (d, J=16.5 Hz, 1H), 6.45 (t, J=75.4 Hz,1H), 6.63 (d, J=16.5 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.66 (dd, J=8.5,1.5 Hz, 1H), 7.80 (br s, 1H), 7.86 (d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz,1H). LCMS (m/z) 616.2 [M+H], Tr=2.58 min.

Example 56 Compound 56

Compound 56a. (R)-2-Methyl-2-trifluoromethoxymethyl-but-3-enoic acid

Zinc bis(trifluoromethylsulfonyl)imide (750 mg, 1.2 mmol) and(1-(trifluoromethyl)-1,2-benziodoxol-3(1H)-one (758 mg, 2.4 mmol) wereadded to a stirred solution of (R)-2-hydroxymethyl-2-methyl-but-3-enoicacid methyl ester (288 mg, 2.0 mmol) in dry deuterochloroform (4 mL)under nitrogen. The reaction mixture was stirred at room temperatureunder nitrogen in a sealed flask for 5 days. The resulting suspensionwas diluted with dichloromethane and the supernatant was separated andevaporated. Dichloromethane (5 mL) was added to the residue and thesupernatant solution was purified by silica gel chromatography using agradient of pentane/diethyl ether 10:1 to 5:1 to afford crude(R)-2-methyl-2-trifluoromethoxymethyl-but-3-enoic acid methyl ester (98mg) as a clear oil which was used in the next step.

A solution of crude (R)-2-methyl-2-trifluoromethoxymethyl-but-3-enoicacid methyl ester (98 mg) in tetrahydrofuran (4 mL) was stirred at 5° C.under nitrogen. A solution of lithium hydroxide monohydrate (63 mg, 1.5mmol) in water (1 mL) was added and the reaction mixture was stirred at5° C. for 1 h and then at room temperature for 3 days. Water was addedand the mixture was acidified to pH 2 with hydrochloric acid (2 M) andthe mixture was extracted with diethyl ether. The organic extracts werecombined and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated to afford thecrude title compound (95 mg) as a clear oil which was used in the nextstep. LCMS (m/z) 197.1 [M−H], Tr=2.16 min.

Compound 56

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (212 mg, 0.4 mmol),crude (R)-2-methyl-2-trifluoromethoxymethyl-but-3-enoic acid (95 mg),tri(o-tolyl)phosphine (24 mg, 0.08 mmol) and triethylamine (121 mg, 0.17mL, 1.2 mmol) in 1,4-dioxane (10 mL) was stirred at 50° C. undernitrogen. Trs(dibenzylideneacetone)dipalladium(0) (36 mg, 0.04 mmol) wasadded and the reaction mixture was heated at reflux for 40 minutes. Thereaction mixture was cooled to room temperature and the mixture wasfiltered through a hydrophobic frit and the filtrate was evaporated toafford crude(E)-(R)-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2-methyl-2-trifluoromethoxymethyl-but-3-enoicacid which was used in the next step. LCMS (m/z) 652.1 [M+H], Tr=2.17min.

A mixture of powdered 4 Å molecular sieves (200 mg),4-dimethylaminopyridine (195 mg, 1.6 mmol), and 2-methyl-6-nitrobenzoicanhydride (275 mg, 0.8 mmol) in dichloromethane (200 mL) was stirred atroom temperature under nitrogen. A solution of crude(E)-(R)-4-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-2-methyl-2-trifluoromethoxymethyl-but-3-enoicacid in dichloromethane (5 mL) was added via syringe pump over 4 h andthe reaction mixture was stirred at room temperature for 18 h. Thereaction mixture was filtered through Celite and the filtrate wasevaporated to a volume of approximately 50 mL. The solution was washedwith ice-cold saturated ammonium chloride solution, ice-cold saturatedsodium hydrogen carbonate solution and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 1:2 to 0:1 then by reverse phase preparativeHPLC eluting with a gradient of acetonitrile/water 1:4 to 1:0 to affordthe title compound (4.3 mg, 0.3% over 4 steps) as a white solid. ¹H NMR(300 MHz, CD₃OD) δ1.01 (d, J=6.7 Hz, 3H), 1.07 (d, J=6.7 Hz, 3H), 1.60(d, J=6.7 Hz, 3H), 1.63 (s, 3H), 1.66 (d, J=7.1 Hz, 3H), 1.66-1.70 (m,2H), 1.93-1.97 (m, 1H), 2.14-2.27 (m, 2H), 2.64-2.73 (m, 1H), 3.58-3.67(m, 1H), 4.10 (d, J=9.1 Hz, 1H), 4.37-4.42 (m, 1H), 4.45 (d, J=9.1 Hz,1H), 5.03 (d, J=12.0 Hz, 1H), 5.09 (q, J=6.7 Hz, 1H), 5.24 (d, J=8.9 Hz,1H), 5.81 (q, J=7.1 Hz, 1H), 6.23 (d, J=16.5 Hz, 1H), 6.64 (d, J=16.5Hz, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.67 (dd, J=8.5, 1.5 Hz, 1H), 7.82 (brs, 1H), 7.87 (d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 634.1[M+H], Tr=2.85 min.

Example 57 Compound 57

Compound 57a. 6-Nitromethyl-[1,4]dioxepan-6-ol

To magnesium sulfate (2.05 g, 17.0 mmol) under nitrogen was addednitromethane (4 mL) and the mixture was vigorously stirred. A solutionof [1,4]ioxepan-6-one (899 mg, 7.74 mmol, prepared as described in PCTInt. Appl. 2010139717) in nitromethane (2 mL) was added and the mixturewas stirred at room temperature for 5 minutes. A solution of2,8,9-triisopropyl-2,5,8,9-tetraaza-1-phosphabicyclo[3,3,3]undecane (231mg, 250 μL, 0.77 mmol) in nitromethane (2 mL) was added and the mixturewas heated at 40° C. for 19 h. The cooled reaction mixture was directlyloaded on silica and purified by silica gel chromatography using agradient of pentane/diethyl ether 1:1 to 1:3 to give the title compound(798 mg, 58%). as a yellow oil. ¹H NMR (300 MHz, CDCl₃) δ3.27 (s, 1H),3.77-3.93 (m, 4H), 3.85 (s, 4H), 4.58 (s, 2H).

Compound 57b. 6-Nitromethylene-[1,4]dioxepane

A stirred mixture of 6-nitromethyl-[1,4]dioxepan-6-ol (0.74 g, 4.18mmol) and triethylamine (1.86 g, 2.56 mL, 18.4 mmol) in dichloromethane(20 mL), under nitrogen, was cooled to −78° C. and methanesulfonylchloride (1.43 g, 970 μL, 12.5 mmol) was added dropwise. The reactionmixture was stirred for 30 minutes and then quenched with saturatedammonium chloride solution. The mixture was removed from the coolingbath and allowed to warm to ambient temperature. The organic layer wasseparated and the aqueous layer was extracted with dichloromethane. Thecombined organic extracts were dried over anhydrous sodium sulfate,filtered and evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 3:1 to 1:1 togive the title compound (512 mg, 77%) as a yellow oil. ¹H NMR (300 MHz,CDCl₃) δ3.81-3.90 (m, 4H), 4.36 (d, J=0.5 Hz, 2H), 5.04 (d, J=2.5 Hz,2H), 7.02 (t, J=2.5 Hz, 1H). LCMS (m/z) 182.1.0 [M+Na], Tr=1.04 min.

Compound 57c. 6-Nitromethyl-6-vinyl-[1,4]dioxepane

A mixture of toluene (13 mL) and water (3.25 mL) was deoxygenated bybubbling nitrogen through for 20 minutes. It was then added to6-nitromethylene-[1,4]dioxepane (512 mg, 3.22 mmol).2,2′-Bis(diphenylphosphino)-1,1′-binaphthalene (218 mg, 0.35 mmol) andpotassium vinyltrifluoroborate (1.73 g, 12.9 mmol) were added and themixture was deoxygenated by bubbling nitrogen through for 5 minutes.Bis(1,5-cyclooctadiene)rhodium(I) tetrafluoroborate (131 mg, 0.32 mmol)was added and the reaction mixture was stirred and heated at refluxunder nitrogen for 4.5 h. The mixture was allowed to cool and waspartitioned between diethyl ether and water. The aqueous layer wasseparated and extracted with diethyl ether and the combined organicsextracts were dried over anhydrous sodium sulfate, filtered andevaporated. The residue was purified by silica gel chromatography usinga gradient of iso-hexanes/diethyl ether 3:1 to 13:7 to give the titlecompound (155 mg, 26%) as a pale brown oil. ¹H NMR (300 MHz, CDCl₃)δ3.77-3.93 (m, 8H), 4.64 (s, 2H), 5.22 (d, J=17.9 Hz, 1H), 5.30 (d,J=11.2 Hz, 1H), 5.79 (dd, J=17.8, 11.2 Hz, 1H). LCMS (m/z) 210.1 [M+Na],Tr=1.51 min.

Compound 57d. 6-Vinyl-[1,4]dioxepane-6-carboxylic acid

To a stirred solution of 6-nitromethyl-6-vinyl-[1,4]dioxepane (214 mg,1.14 mmol) in N,N-dimethylformamide (5 mL) under nitrogen was addedacetic acid (684 mg, 652 μL, 11.4 mmol) and sodium nitrite (237 mg, 3.43mmol). The mixture was heated at 40° C. for 23 h and allowed to cool.The mixture was acidified to pH 1 with hydrochloric acid (2 M) and theaqueous layer was extracted with diethyl ether (3×). The combinedorganic extracts were washed with water, dried over anhydrous magnesiumsulfate, filtered, evaporated. Residual N,N-dimethylformamide wasazeotroped off with toluene to give the title compound (79 mg, 40%) as ayellow gum. The original aqueous layer was saturated with sodiumchloride and extracted with diethyl ether (3×) and the combined organicextracts were washed with water, dried over anhydrous magnesium sulfate,filtered and evaporated. Residual N,N-dimethylformamide was azeotropedoff with toluene to give the title compound (22 mg, 11%) as a yellowgum. The two gums were finally combined (101 mg, 51%). ¹H NMR (300 MHz,CDC₃) δ3.76-3.93 (m, 4H), 3.88 (d, J=12.5 Hz, 2H), 4.35 (d, J=12.7 Hz,2H), 5.26 (d, J=17.8 Hz, 1H), 5.27 (d, J=10.9 Hz, 1H), 5.79 (dd, J=17.4,10.9 Hz, 1H). LCMS (m/z) 195.2 [M+Na], Tr=0.96 min.

Compound 57e.6-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,4]dioxepane-6-carboxylicacid

A mixture of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (68 mg, 0.128 mmol),6-vinyl-[1,4]dioxepane-6-carboxylic acid (22 mg, 0.128 mmol),triethylamine (39 mg, 54 μL, 0.384 mmol) and tri(o-tolyl)phosphine (8mg, 0.026 mmol) in 1,4-dioxane (2 mL) was deoxygenated by bubblingnitrogen through for 5 minutes and then warmed to 50° C. under nitrogenwith stirring. Tris(dibenzylideneacetone)dipalladium(0) (12 mg, 0.013mmol) was added and the mixture was refluxed for 40 minutes and thenallowed to cool. The suspension was filtered and the filtrate wasevaporated to give crude title compound which was used directly in thenext stage. LCMS (m/z) 626.2 [M+H], Tr=1.67 min.

Compound 57

To a stirred mixture of powdered 4 Å molecular sieves (˜1 g),2-methyl-6-nitrobenzoic anhydride (88 mg, 0.256 mmol) and4-(dimethylamino)-pyridine (63 mg, 0.512 mmol) in dichloromethane (38mL), under nitrogen, was added a solution of crude6-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,4]dioxepane-6-carboxylicacid (0.128 mmol) in dichloromethane (5 mL) over 2.5 h via syringe pump.The flask originally containing the crude acid was washed withdichloromethane (1 mL) and added to the reaction mixture over ca. 10minutes. The reaction mixture was stirred for 1 h and then filteredthrough Celite and washed successively with ice-cold saturated ammoniumchloride solution, ice-cold saturated sodium bicarbonate solution,saturated brine and then dried over anhydrous sodium sulfate, filteredand evaporated. The residue was purified by reverse phase preparativeHPLC using a gradient of acetonitrile/water 1:9 to 7:3 containing 0.1%formic acid, to give, after trituration with diethyl ether, the titlecompound (6.5 mg, 8% over 2 steps) as a white solid. ¹H NMR (300 MHz,CD₃OD) δ1.04 (d, J=6.7 Hz, 3H), 1.08 (d, J=6.7 Hz, 3H), 1.53-1.73 (m,2H), 1.60 (d, J=6.7 Hz, 3H), 1.66 (d, J=7.4 Hz, 3H), 1.90-2.00 (m, 1H),2.10-2.28 (m, 2H), 2.63-2.75 (m, 1H), 3.57-3.65 (m, 1H), 3.76-3.91 (m,5H), 4.13 (d, J=12.5 Hz, 1H), 4.38 (d, J=12.7 Hz, 1H), 4.38-4.46 (m,1H), 4.53 (d, J=12.7 Hz, 1H), 5.07 (q, J=6.7 Hz, 1H), 5.35 (d, J=9.2 Hz,1H), 5.76-5.86 (m, 1H), 6.30 (d, J=16.5 Hz, 1H), 6.49 (d, J=16.5 Hz,1H), 7.46 (d, J=8.5 Hz, 1H), 7.68-7.73 (m, 2H), 7.85 (d, J=8.9 Hz, 1H),8.19 (br s, 1H), 8.25 (d, J=8.5 Hz, 1H). LCMS (m/z) 608.2 [M+H], Tr=2.12min.

Example 58 Compound 58

Compound 58a.(S)-4-Isopropyl-3-((S)-2-methoxymethyl-2-methyl-but-3-enoyl)-oxazolidin-2-one

To a solution of(S)-4-isopropyl-3-((E)-2-methyl-but-2-enoyl)-oxazolidin-2-one (preparedaccording to the method reported in Example 29c but using the oppositeenantiomer of the oxazolidinone), (4.44 g, 21 mmol), in anhydroustoluene (40 mL) under nitrogen at −78° C. was added dropwise a solutionof Sodium bis(trimethylsily)amide in toluene (0.6 M, 54 mL, 32 mmol).When addition was completed, the reaction mixture was left to stir at−78° C. for another minutes. Chloromethyl methyl ether (3.38 g, 42 mmol)was slowly added. Stirring was maintained at −60° C. for 30 minutes andthen for 1 h as the temperature was allowed to rise to 0° C. Saturatedammonium chloride solution (230 mL) was added and the mixture was leftto stir at room temperature for 10 minutes. The aqueous layer wasextracted with ethyl acetate (2×) and the combined organic layers werewashed with brine, dried over anhydrous sodium sulfate and filtered. Thefiltrate was evaporated. The crude residue was purified by silica gelchromatography using a Biotage KP-Sil 100 g cartridge eluting with agradient of iso-hexanes/ethyl acetate 1:0 to 9:1 to give the titleproduct (2.55 g, 48%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃)δ0.80-1.00 (m, 6H), 1.50 (s, 3H), 2.25-2.45 (m, 1H), 3.35 (s, 3H), 3.46(d, J=8.7 Hz, 1H), 4.16-4.23 (m, 2H), 4.28 (app t, J=8.2 Hz, 1H),4.45-4.60 (m, 1H), 5.01 (d, J=17.6 Hz, 1H), 5.12 (d, J=10.7 Hz, 1H),6.19 (dd, J=17.6, 10.7 Hz, 1H). LCMS (m/z) 255.3 [M+H], Tr=2.58 min.

Compound 58b. (S)-2-Methoxymethyl-2-methyl-but-3-enoic acid

To a stirred solution of(S)-4-isopropyl-3-((R)-2-methoxymethyl-2-methyl-but-3-enoyl)-oxazolidin-2-one(1.5 g, 5.87 mmol) in tetrahydrofuran (30 mL) and water (15 mL) at 0°C., was added hydrogen peroxide (30% solution, 3 mL, 29.35 mmol) andlithium hydroxide monohydrate (0.59 g, 11.75 mmol). The reaction mixturewas stirred at 0° C. for 3 h then was treated with saturated sodiummetabisulfite and the pH of the solution was adjusted to 1 with dropwiseaddition of hydrochloric acid (10 M). The aqueous layer was extractedwith ethyl acetate (2×) and the combined organic layers were washed withbrine, dried over anhydrous sodium sulfate and filtered. The filtratewas evaporated. The crude residue was purified by silica gelchromatography using a Biotage KP-Sil 50 g cartridge, eluting with agradient of iso-hexanes/ethyl acetate 1:0 to 3:1 to give the titleproduct (0.546 g, 65%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ1.36(s, 3H), 3.42 (s, 3H), 3.45 (d, J=8.9 Hz, 1H), 3.62 (d, J=8.9 Hz, 1H),5.20-5.30 (m, 2H), 6.01 (dd, J=17.6, 10.7 Hz, 1H), 9.80-11.80 (br. s,1H).

Compound 58c.(E)-(S)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2-methoxymethyl-2-methyl-but-3-enoicacid

To a vial charged with [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamicacid tert-butyl ester (706 mg, 2 mmol),(S)-2-methoxymethyl-2-methyl-but-3-enoic acid (290 mg, 2 mmol),tri-(o-tolyl)phosphine (610 mg, 2 mmol) andN,N-dicyclohexyl-methyl-amine (0.85 mL, 4 mmol), was added acetonitrile(2 mL) and 1,4-dioxane (6 mL). The vial was purged with nitrogen.Palladium(II) acetate (450 mg, 2 mmol) was added. The vial was sealedand irradiated by microwave at 130° C. for 1 h. The mixture was thentreated with saturated solution of potassium hydrogen sulfate. Theaqueous layer was extracted with ethyl acetate (2×) and the combinedorganic layers were washed with brine, dried over anhydrous sodiumsulfate and filtered. The filtrate was evaporated. The crude residue waspurified by silica gel chromatography using a Biotage KP-Sil 100 gcartridge eluting with a gradient of ethyl acetate/methanol 1:0 to 41:9to give the title product (0.176 g, 21%) as an amber oil. ¹H NMR (300MHz, CDCl₃) δ1.45 (s, 9H), 1.54 (s, 3H), 1.57 (d, J=6.5 Hz, 3H), 3.47(s, 3H), 3.62 (d, J=8.7 Hz, 1H), 3.77 (d, J=8.7 Hz, 1H), 4.90-5.10 (m,1H), 6.60-6.85 (m, 2H), 7.34 (d, J=8.4 Hz, 1H), 7.61 (d, J=8.4 Hz, 1H),7.75 (d, J=8.4 Hz, 1H), 8.06-8.17 (m, 2H). LCMS (m/z) 415.3 [M+H],Tr=2.11 min.

Compound 58d.(S)-1-[(S)-2-((S)-2-{(E)-(S)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2-methoxymethyl-2-methyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

To a solution of(E)-(S)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2-methoxymethyl-2-methyl-but-3-enoicacid (176 mg, 0.34 mmol) in dichloromethane (2 mL), was added(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (205 mg, 0.47 mmol),4-dimethylaminopyridine (116 mg, 0.95 mmol), N,N-diisopropylethylamine(123 mg, 0.95 mmol) and 2-methyl-6-nitrobenzoic anhydride (164 mg, 0.47mmol). The reaction mixture was stirred at room temperature for 18 h,was then treated with saturated solution of sodium hydrogen carbonate,extracted with dichloromethane (2×) and the combined organic layers werewashed with brine, dried over anhydrous sodium sulfate and filtered. Thefiltrate was evaporated. The crude residue was purified by silica gelchromatography using a Biotage KP-Sil 50 g cartridge eluting with agradient of iso-hexanes/ethyl acetate 9:1 to 0:1 to give the titleproduct (0.22 g, 78%) as a yellow solid. ¹H NMR (300 MHz, CDCl₃)δ0.85-1.10 (m, 6H), 1.20-1.70 (m, 23H), 2.05-2.20 (m, 1H), 2.30-2.45 (m,1H), 3.44 (s, 3H), 3.60-3.95 (m, 4H), 4.15-4.32 (m, 1H), 4.68-4.70 (m,1H), 4.90-5.05 (m, 2H), 5.20-5.45 (m, 1H), 6.10-6.28 (m, 1H), 6.60-6.85(m, 2H), 7.10-7.20 (m, 1H), 7.28 (d, J=8.4 Hz, 1H), 7.66-7.74 (m, 2H),8.00 (d, J=4.0 Hz, 1H), 8.05 (d, J=8.4 Hz, 1H). LCMS (m/z) 828.5 [M+H],Tr=3.48 min.

Compound 58

To a solution of(S)-1-[(S)-2-((S)-2-{(E)-(S)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2-methoxymethyl-2-methyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (0.22 g, 0.265 mmol) in tetrahydrofuran(3 mL), was added zinc powder (190 mg, 2.92 mmol) and a solution ofammonium acetate (153 mg, 1.99 mmol) in water (1.5 mL). The resultingmixture was stirred for 24 h. The mixture was filtered through a pad ofCelite and the filtrate was washed with saturated potassium hydrogensulfate. The aqueous layer was extracted with ethyl acetate. Thecombined organic layers were washed with brine, dried over anhydroussodium sulfate and filtered. The filtrate was evaporated and thenco-evaporated with toluene to give an amber oil which was treated withhydrogen chloride in 1,4-dioxane (4 M, 4 mL) After stirring for 1 hvolatiles were evaporated to give a crude solid which was dissolved inanhydrous dichloromethane (20 mL) and anhydrous N,N-dimethylformamide (1mL). N,N-Diisopropylethylamine (66 mg, 0.515 mmol) and2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (59 mg, 0.155 mmol) were added. Themixture was stirred for 1.5 h then water was added and the organic layerwas filtered through a phase separator frit. The organic filtrate wasevaporated and then co-evaporated with toluene to give an amber oil. Thecrude residue was purified by silica gel chromatography using a BiotageKP-Sil 10 g cartridge eluting with a gradient of iso-hexanes/ethylacetate 9:1 to 0:1 then by reverse phase preparative HPLC eluting with agradient of water/acetonitrile 13:7 to 13:12 over 30 minutes to give thetitle compound.

Compound 58 (2.83 mg, 4.8%) as white solid. ¹H NMR (300 MHz, CD₃OD)δ1.02 (d, J=6.7 Hz, 3H), 1.08 (d, J=6.7 Hz, 3H), 1.48 (s, 3H), 1.50-1.80(m, 7H), 1.91-2.03 (m, 1H), 2.11-2.22 (m, 1H), 2.24-2.34 (m, 1H),2.65-2.78 (m, 1H), 3.39 (s, 3H), 3.43 (d, J=8.5 Hz, 1H), 3.54-3.67 (m,1H), 3.99 (d, J=8.5 Hz, 1H), 4.40-4.50 (m, 1H), 5.02-5.20 (m, 2H), 5.30(d, J=8.9 Hz, 1H), 5.82 (q, J=7.1 Hz, 1H), 6.31 (d, J=16.5 Hz, 1H), 6.64(d, J=16.5 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.64 (dd, J=8.2, 1.5 Hz,1H), 7.79 (s, 1H), 7.85 (d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS(m/z) 580.2 [M+H], Tr=2.45 min.

Example 59 Compound 59

Compound 59a

To a solution of L-valine-d₈ (CAS Number: 35045-72-8; C/DIN IsotopesInc.) (1 g, 8 mmol) in 1M sulfuric acid (16 ml, 1M aqueous solution),cooled to 0° C., was added a solution of sodium nitrite (1.1 g, 16 mmol)in water (8 ml). The temperature was maintained below 5° C. during theaddition, and the mixture was stirred at such overnight. The solutionwas then saturated with ammonium sulfate, extracted with diethyl ether(3×25 ml), dried over sodium sulfate and evaporated under reducedpressure giving the title compound (0.68 g, 67%) as a colorless oil thatcrystallized on standing. ¹H NMR (400 MHz, CD₃OD): 6 no signal. LCMS(m/z) 125.0 [M−H], Tr=0.65 min.

Compound 59b

A solution of 1d (866 mg, 2 mmol) in dichloromethane (30 mL) was cooledin an ice water bath. Trimethylsilyl trifluoromethanesulfonate (667 mg,3 mmol) was added dropwise at 0° C. under argon, and the resultingsolution was stirred at room temperature for 30 minutes. The reactionmixture was evaporated to dryness and the resulting crude residue (LCMS(m/z) 332.2/334.3 [M+H]′ Tr=2.06 min) was dissolved in anhydrousdichloromethane (20 mL) and cooled to 0° C. To this was added 59a (252mg, 2.2 mmol), N,N-diisopropylethylamine (0.86 mL, 5 mmol), and(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(1062 mg, 2.4 mmol). The reaction was allowed to warm to roomtemperature and stirred for 8 hours. The mixture was evaporated todryness and the residue was dissolved in ethyl acetate (100 mL) andwashed with 10% citric acid (100 mL), saturated NaHCO₃ (100 mL), andbrine (100 mL). The organic layer was dried over MgSO₄, one volumeequivalent of hexane was added and this solution was filtered through a5 cm layer of silica gel, silica gel layer was washed with 50 mL ofethyl acetate/hexane mixture (1/1). The desired product was washed outwith ethyl acetate (100 mL), concentrated under reduced pressure andco-distilled with dichloromethane. 59b (878 mg, quantitative yield) wasisolated after drying under high vacuum for one day. ¹H NMR (400 MHz,CD₃OD) δ5.50-5.42 (m, 1H), 5.03 (d, J=12.1 Hz, 1H), 4.85 (d, J=11.9 Hz,1H), 4.16-4.10 (m, 1H), 3.88-3.85 (m, 1H), 3.75-3.41 (m, 2H), 2.16-2.07(m, 1H), 2.00-1.83 (m, 2H), 1.81-1.69 (m, 1H), 1.32 (d, J=6.8 Hz, 3H).LCMS (m/z) 440.1/442.1 [M+H]′ Tr=2.32 min.

Compound 59c

Into an oven-dried, argon purged flask were placed2-methyl-6-nitrobenzoic anhydride (344 mg, 1 mmol),4-dimethylaminopyridine (128 mg, 1.05 mmol), Boc-protected quinolinecarboxylic acid (192 mg, 0.50 mmol), and anhydrous dichloromethane (20mL). Into the resulting solution was added N,N-diisopropylethylamine(0.26 mL, 1.50 mmol) and this reaction mixture was stirred at roomtemperature for 10 minutes. 59b (331 mg, 0.75 mmol) was added dropwisevia syringe as a solution in anhydrous dichloromethane (10 mL). Afterstirring for 12 hours at room temperature, the reaction mixture wastransferred to a separatory funnel and washed with water (20 mL, 10 mLof brine was added to support the separation). The aqueous phase wasextracted with dichloromethane (20 mL). Combined organic extracts werewashed with brine (20 mL) and dried over magnesium sulfate, filtered andconcentrated under reduced pressure. The residue was purified by silicagel chromatography (gradient from 0-40% ethyl acetate+methanol (4/1) iniso-hexanes) to afford 59c (391 mg, 97%) as a white solid afterevaporation. ¹H NMR (400 MHz, CD₃OD) δ 8.28 (d, J=8.5 Hz, 1H), 7.97 (s,1H), 7.87 (d, J=8.5 Hz, 1H), 7.81 (d, J=8.5 Hz, 1H), 7.52 (d, J=8.5 Hz,1H), 6.90-6.71 (m, 2H), 5.49-5.34 (m, 2H), 5.50-4.98 (m, 2H), 4.87-4.79(m, 2H), 4.17-4.10 (m, 1H), 4.90-4.80 (m, 2H), 2.15-2.06 (m, 1H),1.99-1.85 (m, 2H), 1.82-1.72 (m, 1H), 1.72-1.62 (m, 1H), 1.58 (d, J=5.8Hz, 3H), 1.52 (d, J=7.1 Hz, 3H), 1.47 (s, 9H), 1.32 (dd, J=13.9, 6.9 Hz,6H). LCMS (m/z) 806.2/808.1 [M+H]′ Tr=2.82 min.

Compound 59

To 59c (390 mg, 0.48 mmol) in tetrahydrofuran (20 mL) was added asolution of lithium hydroxide (13 mg, 0.53 mmol) in water (10 mL). Afterstirring at room temperature for 2 hour, 1M hydrochloric acid was added(0.55 mL of 1M solution in water, 0.55 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (10 mL, 40 mmol) at room temperature under argon for 4hours. Reaction mixture was concentrated under reduced pressure. Thisresidue was dissolved in N,N-dimethylformamide (5 mL) and the obtainedsolution was added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (276 mg, 0.73 mmol),N,N-diisopropylethylamine (312 mg, 2.42 mmol) and dichloromethane (200mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (100 ml) and brine (100 mL), dried over magnesium sulfate,filtered and concentrated under reduced pressure. The residue waspurified by silica gel chromatography (gradient from 0-40% ethylacetate+methanol (4/1) in iso-hexanes) to afford 59 (137 mg, 51%) as awhite solid after evaporation. ¹H NMR (400 MHz, CD₃OD) δ 8.27 (d, J=8.5Hz, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.80 (s, 1H), 7.64 (dd, J=8.5, 1.6 Hz,1H), 7.46 (d, J=8.5 Hz, 1H), 6.56 (d, J=16.3 Hz, 1H), 6.38 (d, J=16.3Hz, 1H), 5.82 (q, J=7.3 Hz, 1H), 5.11 (q, J=6.7 Hz, 1H), 4.45 (dd,J=13.0, 3.5 Hz, 1H), 3.62 (dd, J=11.8, 2.8 Hz, 1H), 2.72 (td, J=13.0,3.3 Hz, 1H), 2.40-2.65 (m, 1H), 2.02-1.94 (m, 1H), 1.77-1.68 (m, 2H),1.65 (d, J=7.3 Hz, 3H), 1.60 (d, J=6.7 Hz, 3H), 1.55 (s, 3H), 1.44 (s,3H). LCMS (m/z) 558.4 [M+H]′ Tr=2.41.

Example 60 Compound 60

Compound 60a

To a solution of (2S,3R)-2-amino-3-methoxybutanoic acid (CAS Number:4144-02-9; Ark Pharm, Inc.) (1 g, 7.5 mmol) in 1M sulfuric acid (15 ml,1M aqueous solution), cooled to 0° C., was added a solution of sodiumnitrite (1.0 g, 15 mmol) in water (8 ml). The temperature was maintainedbelow 50° C. during the addition, and the mixture was stirred at suchovernight. The solution was then saturated with ammonium sulfate,extracted with diethyl ether (5×25 ml), dried over sodium sulfate andevaporated under reduced pressure giving 60a (0.27 g, 67%) as acolorless oil that crystallized on standing. ¹H NMR (400 MHz, CDCl₃):δ4.14 (d, J=3.6 Hz, 1H), 3.81 (qd, J=6.4, 3.6 Hz, 1H), 3.41 (s, 3H),1.25 (d, J=6.4 Hz, 3H). LCMS (m/z) 132.9 [M−H], Tr=0.39 min.

Compound 60b

A solution of 1d (517 mg, 1.19 mmol) in dichloromethane (20 mL) wascooled in an ice water bath. Trimethylsilyl trifluoromethanesulfonate(398 mg, 1.79 mmol) was added dropwise at 0° C. under argon, and theresulting solution was stirred at room temperature for 30 minutes. Thereaction mixture was evaporated to dryness and the resulting cruderesidue (LCMS (m/z) 332.2/334.3 [M+H]′ Tr=2.06 min) was dissolved inanhydrous dichloromethane (20 mL) and cooled to 0° C. To this was added60a (192 mg, 1.43 mmol), N,N-diisopropylethylamine (0.51 mL, 2.98 mmol),and (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (633 mg, 1.43 mmol). The reaction was allowed towarm to room temperature and stirred for 8 hours. The mixture wasevaporated to dryness and the residue was dissolved in ethyl acetate(100 mL) and washed with 10% citric acid (100 mL), saturated NaHCO₃ (100mL), and brine (100 mL). The organic layer was dried over MgSO₄, onevolume equivalent of hexane was added and this solution was filteredthrough a 5 cm layer of silica gel, silica gel layer was washed with 50mL of ethyl acetate/hexane mixture (1/1). The desired product was washedout with ethyl acetate (100 mL), concentrated under reduced pressure andco-distilled with dichloromethane. The 60b (537 mg, quantitative yield)was isolated after drying under high vacuum for one day. ¹H NMR (400MHz, CD₃OD): δ5.53 (q, J=6.9 Hz, 1H), 5.10 (d, J=12.1 Hz, 1H), 4.92 (d,J=12.2 Hz, 1H), 3.96 (d, J=2.6 Hz, 1H), 3.94 (dd, J=7.1, 4.7 Hz, 1H),3.86 (qd, J=6.4, 2.5 Hz, 1H), 3.81-3.60 (m, 2H), 3.42 (s, 3H), 2.34-2.14(m, 1H), 2.09-1.90 (m, 2H), 1.87-1.76 (m, 1H), 1.41 (d, J=6.9 Hz, 3H),1.32 (d, J=6.4 Hz, 3H). LCMS (m/z) 448.1/450.0[M+H]′ Tr=2.11 min.

Compound 60c

Into an oven-dried, argon purged flask were placed2-methyl-6-nitrobenzoic anhydride (172 mg, 0.5 mmol),4-dimethylaminopyridine (64 mg, 0.50 mmol), N-Boc protected(R,E)-4-(2-(1-(tert-butoxycarbonylamino)ethyl)quinolin-7-yl)-2,2dimethylbut-3-enoic acid (96 mg, 0.25 mmol), and anhydrousdichloromethane (10 mL). Into the resulting solution was addedN,N-diisopropylethylamine (0.13 mL, 0.75 mmol) and this reaction mixturewas stirred at room temperature for 10 minutes. 60b (168 mg, 0.38 mmol)was added dropwise via syringe as a solution in anhydrousdichloromethane (10 mL). After stirring for 12 hours at roomtemperature, the reaction mixture was transferred to a separatory funneland washed with water (20 mL, 10 mL of brine was added to support theseparation). The aqueous phase was extracted with dichloromethane (20mL). Combined organic extracts were washed with brine (20 mL) and driedover magnesium sulfate, filtered and concentrated under reducedpressure. The residue was purified by silica gel chromatography(gradient from 0-40% ethyl acetate+methanol (4/1) in iso-hexanes) toafford 60c (149 mg, 73%) as a white solid after evaporation. ¹H NMR (400MHz, CD₃OD): δ8.35 (d, J=8.6 Hz, 1H), 8.07-8.01 (m, 1H), 7.93 (d, J=8.5Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 6.98-6.77 (m,2H), 5.48 (q, J=6.5, 6.1 Hz, 1H), 5.20 (d, J=4.0 Hz, 1H), 5.06 (d,J=12.1 Hz, 1H), 4.88 (d, J=12.1 Hz, 1H), 4.25-4.15 (m, 1H), 3.99-3.85(m, 2H), 3.80-3.55 (m, 2H), 3.46 (s, 3H), 2.20-2.09 (m, 1H), 2.07-1.94(m, 1H), 1.92-1.80 (m, 1H), 1.80-1.71 (m, 1H), 1.68-1.64 (m, 6H), 1.59(d, J=7.1 Hz, 3H), 1.54 (s, 9H), 1.37 (d, J=7.0 Hz, 3H), 1.29 (d, J=6.4Hz, 3H). LCMS (m/z) 814.1/816.1 [M+H]′ Tr=2.64 min.

Compound 60

To 60c (139 mg, 0.17 mmol) in tetrahydrofuran (10 mL) was added asolution of lithium hydroxide (4.5 mg, 0.19 mmol) in water (5 mL). Afterstirring at room temperature for 2 hour, 1M hydrochloric acid was added(0.20 mL of 1M solution in water, 0.20 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (5 mL, 20 mmol) at room temperature under argon for 4 hours.Reaction mixture was concentrated under reduced pressure. This residuewas dissolved in N,N-dimethylformamide (5 mL) and the obtained solutionwas added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (98 mg, 0.26 mmol),N,N-diisopropylethylamine (110 mg, 0.86 mmol) and dichloromethane (100mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (50 ml) and brine (50 mL), dried over magnesium sulfate, filteredand concentrated under reduced pressure. The residue was purified bysilica gel chromatography (gradient from 0-60% ethyl acetate+methanol(4/1) in iso-hexanes) to afford 60 (32 mg, 33%) as a white solid afterevaporation. ¹H NMR (400 MHz, CD₃OD): δ8.24 (d, J=8.5 Hz, 1H), 7.83 (d,J=8.4 Hz, 1H), 7.81 (s, 1H), 7.61 (dd, J=8.4, 1.6 Hz, 1H), 7.43 (d,J=8.5 Hz, 1H), 6.54 (d, J=16.3 Hz, 1H), 6.37 (d, J=16.3 Hz, 1H), 5.78(q, J=7.2 Hz, 1H), 5.44 (d, J=8.2 Hz, 1H), 5.08 (q, J=6.7 Hz, 1H), 4.41(dd, J=13.5, 3.4 Hz, 1H), 3.77-3.63 (m, 1H), 3.59 (dd, J=11.8, 2.9 Hz,1H), 3.41 (s, 3H), 2.68 (td, J=12.9, 3.1 Hz, 1H), 2.31-2.24 (m, 1H),1.99-1.91 (m, 1H), 1.72-1.64 (m, 2H), 1.61 (d, J=7.2 Hz, 3H), 1.57 (d,J=6.7 Hz, 3H), 1.53 (s, 3H), 1.42 (s, 3H), 1.26 (d, J=6.4 Hz, 3H). LCMS(m/z) 566.3 [M+H]′ Tr=2.42 min.

Example 61 Compound 61

Compound 61a

To 1d (500.3 mg, 1.16 mmol) was added an HCl solution (1.2 mL, 4 M indioxane, 1 M). After stirring for 18 hours, the reaction wasconcentrated in vacuo to afford the free dipetide as an amorphous, paleyellow solid.

To L-(+)-lactic acid (252.3 mg, 2.80 mmol) in DMF (10 mL, 0.3 M) wasadded TMSCl (0.35 mL, 2.77 mmol), followed by i-Pr₂NEt (1.0 mL, 5.74mmol). After 3 hours, HATU (1.08 g, 2.84 mmol) was added. After 10minutes, the free dipeptide was added in DMF (11.5 mL, 1 M). After 60minutes, the reaction was concentrated in vacuo to 5 mL, diluted withEtOAc (10 mL) and washed with water (10 mL). The aqueous was extractedwith EtOAc (2×10 mL) and the combined organics were washed with 5% aq.LiCl (5×20 mL) and brine (20 mL), dried over MgSO₄, and concentrated invacuo. The compound was purified via column chromatography (0-100%EtOAc/hexane) to afford 61a (346.6 g, 74%) as a white foam. ¹H NMR (400MHz, CDCl₃): δ7.22 (d, J=8.1 Hz, 1H), 5.36 (p, J=6.9 Hz, 1H), 4.95 (d,J=11.9 Hz, 1H), 4.72 (d, J=11.9 Hz, 1H), 4.34 (d, J=12.1 Hz, 1H), 4.23(q, J=6.8 Hz, 1H), 3.70 (m, 1H), 3.16 (m, 1H), 2.91 (m, 1H), 2.18 (m,1H), 1.94 (m, 1H), 1.72 (m, 2H), 1.49 (dd, J=7.1, 3.4 Hz, 2H), 1.43 (t,J=7.0 Hz, 3H), 1.34 (d, J=6.9 Hz, 3H). LCMS (m/z) 477.85 [M+H].

To the N-Boc(R,E)-4-(2-(1-(tert-butoxycarbonylamino)ethyl)quinolin-7-yl)-2,2dimethylbut-3-enoic acid (150.9 mg, 0.390 mmol), 61a (191.6 mg, 0.468mmol), and DMAP (115.3 mg, 0.936 mmol) in DCM (2.2 mL, 0.1 M) at 0° C.under Ar was added i-Pr₂NEt (0.16 mL, 0.936 mmol) followed by2-methyl-6-nitrobenzoic anhydride (278.6 mg, 0.780 mmol). The reactionwas warmed to rt and, after 60 minutes, was diluted with DCM (10 mL),washed with sat. NH₄Cl_((aq)) (15 mL). The aqueous was extracted withDCM (2×15 mL). The combined organics were washed with H₂O (50 mL), driedover MgSO₄, and concentrated in vacuo. The compound was purified viacolumn chromatography (0-75% EtOAc/hexane) to afford 61b (214.8 mg, 71%)as a yellow oil. ¹H NMR (400 MHz, CDCl₃): δ8.11 (d, J=7.4 Hz, 1H), 8.04(m, 2H), 7.72 (d, J=8.8 Hz, 1H), 7.65 (d, J=8.4 Hz, 1H), 6.70 (dd,J=20.5, 10.7 Hz, 4H), 5.22 (m, 2H), 4.94 (dd, J=11.8, 8.2 Hz, 3H), 4.71(d, J=11.9 Hz, 2H), 4.32 (m, 1H), 3.68 (m, 2H), 2.82 (m, 3H), 2.16 (m,1H), 1.89 (m, 1H), 1.68 (m, 2H), 1.54 (s, 6H), 1.51 (d, J=7.5 Hz, 6H),1.47 (s, 9H), 1.42 (d, J=6.8 Hz, 3H), 1.29 (m, 3H). LCMS (m/z) 770.17[M+H].

Compound 61

To 61b (214.8 mg, 0.279 mmol) in THF (2.8 mL, 0.1 M) at 0° C. was added0.3 M NaOH_((aq)) (0.93 mL, 0.249 mmol). After 40 minutes, the reactionwas quenched with 1 M HCl_((aq)) (0.29 mL) to ˜pH 4. The mixture wasthen diluted with EtOAc (15 mL), washed with water (3×5 mL) and brine(10 mL), dried over MgSO₄, and concentrated in vacuo to afford a yellowfoam (190.1 mg). The crude material was dissolved in dioxane (1 mL) andan HCl solution (3 mL, 4 M in dioxane) was added. After 18 hours, thesuspension was concentrated in vacuo to a yellow solid (175.3 mg). Thecrude material in DMF (5 mL) was diluted with DCM (750 mL, 0.0003 M) andi-Pr₂NEt (0.30 mL, 1.62 mmol) was added, followed by HATU (151.9 mg,0.400 mmol). After 20 minutes, the mixture was washed with 5%LiCl_((aq)) (2×500 mL) and brine (500 mL), dried over MgSO₄, andconcentrated in vacuo to an orange residue. The impurities weretriturated with DCM/tBME to afford 61 (68.0 mg, 47%) as a yellowresidue. ¹H NMR (400 MHz, CDCl₃): δ 8.98 (s, 1H), 8.25 (s, 1H), 8.11 (s,1H), 7.74 (d, J=7.0 Hz, 2H), 7.45 (s, 2H), 6.69 (d, J=15.4 Hz, 2H), 6.43(d, J=16.1 Hz, 1H), 5.80 (m, 1H), 5.41 (m, 1H), 4.57 (d, J=8.9 Hz, 2H),4.15 (s, 1H), 3.77 (m, 1H), 3.66 (m, 4H), 3.55 (m, 2H), 3.08 (m, 2H),2.58 (m, 1H), 1.97 (d, J=14.1 Hz, 1H), 1.56 (m, 6H), 1.44 (m, 6H). LCMS(m/z) 522.23 [M+H]. Rt=4.75 min.

Example 62 Compound 62

Compound 62a

To L-(−)-malic acid (10 g, 74.6 mol) and p-toluenesulfonic acidmonohydrate (172.5 mg, 7.46 mmol) was added 2,2-dimethoxypropane (75 mL,1 M). After 18 hours, water (50 mL) and NaHCO₃ (72.3 mg) were added, andthe mixture was extracted with DCM (3×100 mL). The combined organicswere dried over MgSO₄ and concentrated in vacuo to a yellow oil. Thematerial was purified via crystallization with Et₂O/hexane to afford 62a(11.2 g, 88%) as a white solid. ¹H NMR (400 MHz, CDCl₃): δ4.72 (dd,J=6.6, 3.9 Hz, 1H), 3.00 (dd, J=17.3, 3.9 Hz, 1H), 2.86 (dd, J=17.3, 6.6Hz, 1H), 1.63 (s, 3H), 1.57 (s, 3H).

Compound 62b

To 62a (385.9 g, 2.21 mmol) in DCM (22 mL, 0.1 M) at 0° C. was addedoxalyl chloride (0.23 mL, 2.66 mmol) followed by a drop of DMF. Thereaction was warmed to rt and stirred for 60 minutes. The reaction wasthen concentrated in vacuo to afford pale yellow needles. The crude wasdissolved in toluene (22 mL, 0.1 M) and Pd/BaSO₄ (111.9 mg) was added.The mixture was purged with H₂ and refluxed for 3 hours. The reactionwas then cooled to rt, filtered through celite, rinsed with EtOAc, andconcentrated in vacuo to afford 62b (160 mg, 47%) as a yellow oil. ¹HNMR (400 MHz, CDCl₃): δ9.78 (s, 1H), 4.79 (dd, J=7.0, 3.5 Hz, 1H), 3.09(dd, J=18.3, 3.5 Hz, 1H), 2.92 (dd, J=18.3, 7.0 Hz, 1H), 1.63 (s, 3H),1.58 (s, 3H).

Compound 62c

To 62b (1 g, 5.55 mmol) in DCM (11 mL, 0.5 M) at −15° C. was added DAST(0.88 mL, 6.66 mmol) and the reaction was allowed to warm to rt slowly.After 14 hours, the reaction was slowly quenched with cold sat.NaHCO_(3(aq)) (10 mL), the phases were separated, and the aqueous wasextracted with DCM (2×15 mL). The combined organics were dried overMgSO4 and concentrated in vacuo to afford 62c (956.2 mg, 92%) as ayellow-orange oil. ¹H NMR (400 MHz, CDCl₃): δ6.01 (tdd, J=56.1, 5.4, 4.1Hz, 1H), 4.53 (ddd, J=8.4, 4.0, 0.8 Hz, 1H), 2.52-2.36 (m, 1H),2.32-2.15 (m, 1H), 1.65-1.60 (m, 3H), 1.56-1.53 (m, 3H). ¹⁹F NMR (376MHz, CDCl₃) δ ⁻117.10-⁻119.71 (m).

Compound 62d

To 62c (956.2 mg, 5.31 mmol) in THF (5 mL, 1 M) was added 1 M HCl_((aq))(5 mL, 1 M). After 18 hours, the reaction was saturated with NaCl, andextracted Et₂O (2×20 mL) and THF (15 mL). The combined organics weredried over MgSO₄ and concentrated in vacuo to afford 62d (654.8 mg, 88%)as a clear oil. ¹H NMR (400 MHz, CDCl₃): δ6.06 (tdd, J=56.4, 5.5, 4.2Hz, 1H), 4.48-4.39 (m, 1H), 2.51-2.33 (m, 1H), 2.33-2.12 (m, 1H). ¹⁹FNMR (376 MHz, CDCl₃) δ ⁻117.40-⁻120.28 (m).

Compound 62e

To 1d (2.75 g, 6.37 mmol) in dioxane (2 mL) was added an HCl solution(6.5 mL, 25.5 mmol, 4 M in dioxane). After stirring for 18 hours, thereaction was concentrated in vacuo to afford the free dipetide as anamorphous, pale yellow solid.

To 62d (654.8 mg, 4.67 mmol) in DMF (16 mL, 0.3 M) was added TMSCl (0.59mL, 4.67 mmol), followed by i-Pr₂NEt (2.0 mL, 11.7 mmol). After 2 hours,the free dipetide was added in DMF (6 mL, 1 M) followed by i-Pr₂NEt (4.0mL, 23.4 mmol) and HATU (2.66 g, 7.01 mmol). After 45 minutes, thereaction was concentrated in vacuo to 5 mL, diluted with sat.NaHCO_(3(aq)) (50 mL) and extracted with EtOAc (3×50 mL). The combinedorganics were washed with 5% LiCl_((aq)) (4×50 mL) and brine (100 mL),dried over MgSO₄, and concentrated in vacuo. The compound was purifiedvia column chromatography (50-100% EtOAc/hexane) to afford 62e (1.24 g,59%) as a tan foam. ¹H NMR (400 MHz, CDCl₃): δ6.06 (m, 1H), 5.36 (p,J=7.2 Hz, 1H), 4.95 (d, J=12.0 Hz, 1H), 4.72 (d, J=11.8 Hz, 1H), 4.32(m, 2H), 3.85 (m, 1H), 3.70 (m, 1H), 2.94 (m, 1H), 2.39 (m, 1H), 2.16(m, 2H), 1.94 (m, 1H), 1.74 (m, 2H), 1.34 (d, J=6.9 Hz, 3H). ¹⁹F NMR(376 MHz, CDCl₃) δ −117.56 (dt, J=56.4, 16.5 Hz). LCMS (m/z) 456.03[M+H].

Compound 62f

To N-Boc(R,E)-4-(2-(1-(tert-butoxycarbonylamino)ethyl)quinolin-7-yl)-2,2dimethylbut-3-enoic acid (98.7 mg, 0.220 mmol), 62e (115.3 mg, 0.264mmol), and DMAP (66.1 mg, 0.528 mmol) in DCM (2.2 mL, 0.1 M) under Arwas added i-Pr₂NEt (0.10 mL, 0.528 mmol) followed by2-methyl-6-nitrobenzoic anhydride (115.9 mg, 0.330 mmol). After 60minutes, the reaction was diluted with DCM (10 mL), washed with sat.NH₄Cl_((aq)) (10 mL). The aqueous was extracted with DCM (10 mL). Thecombined organics were washed with H₂O (15 mL), dried over MgSO₄, andconcentrated in vacuo. The compound was purified via columnchromatography (20-100% EtOAc/hexane) to afford 62f (159.0 mg, 89%) asan pale yellow foam. ¹H NMR (400 MHz, CDCl₃): δ8.02 (d, J=15.5 Hz, 1H),7.78-7.60 (m, 3H), 7.18 (s, 1H), 6.71 (m, 2H), 5.93 (tt, J=55.9, 4.4 Hz,1H), 5.37 (dd, J=7.5, 4.3 Hz, 1H), 5.23 (m, 1H), 4.91 (d, J=11.9 Hz,1H), 4.69 (d, J=11.9 Hz, 1H), 4.25 (d, J=9.7 Hz, 1H), 3.76 (d, J=11.0Hz, 1H), 3.64 (m, 1H), 2.84 (m 1H), 2.48 (m, 1H), 2.15 (m, 2H), 1.88(dt, J=3.8, 2.8 Hz, 2H), 1.67 (dq, J=12.4, 6.4 Hz, 6H), 1.55 (d, J=7.0Hz, 9H), 1.44 (s, 12H). ¹⁹F NMR (376 MHz, cdcl₃) δ −115.88 (ddt, J=55.4,33.0, 17.6 Hz). LCMS (m/z) 822.19 [M+H].

Compound 62

To 62f (158.0 mg, 0.194 mmol) in THF (2.0 mL, 0.1 M) at 0° C. was added0.3 M NaOH_((aq)) (0.65 mL, 0.194 mmol). After 20 minutes, the reactionwas quenched with 1 M HCl_((aq)) (0.2 mL) to ˜pH 4. The mixture was thendiluted with EtOAc (20 mL), washed with water (3×5 mL) and brine (5 mL),dried over MgSO₄, and concentrated in vacuo to afford a yellow residue(153.9 mg). The crude material was dissolved in dioxane (1 mL) and anHCl solution (1 mL, 4 M in dioxane) was added. After 2 hours, thesuspension was concentrated in vacuo to a yellow solid (202.5 mg). Thecrude material in DMF (2 mL, 0.1 M) was diluted with DCM (650 mL, 0.0003M) and i-Pr₂NEt (0.17 mL, 0.982 mmol) was added, followed by HATU (89.9mg, 0.232 mmol). After 30 minutes, the mixture was washed with 5% LiCl(2×300 mL), sat. NaHCO₃ (300 mL), and brine (300 mL), dried over MgSO₄,and concentrated in vacuo. The crude material was purified by prep HPLC(Gemini, 30-90% MeCN/H₂O) to afford 62 (39.2 mg, 36%) as an off-whitesolid. ¹H NMR (400 MHz, CDCl₃): δ9.02 (s, 1H), 8.09 (d, J=18.2 Hz, 2H),7.76 (s, 1H), 7.45 (s, 1H), 6.70 (d, J=16.0 Hz, 2H), 6.43 (d, J=7.9 Hz,1H), 6.30 (d, J=15.8 Hz, 1H), 5.99 (d, J=55.2 Hz, 1H), 5.74 (s, 1H),5.48 (s, 1H), 5.23 (s, 1H), 5.11 (s, 1H), 4.92 (s, 1H), 4.56 (d, J=12.9Hz, 1H), 3.55 (m, 2H), 2.58 (t, J=11.7 Hz, 2H), 2.40 (m, 4H), 1.67 (m,6H), 1.58 (m, 5H), 1.49 (m, 9H). ¹⁹F NMR (376 MHz, CDCl₃) δ −117.00(ddt, J=61.8, 56.0, 14.9 Hz). LCMS (m/z) 690.33 [M+H]. Tr=5.178 min.

Example 63 Compound 63

Compound 63a

To (S)-2-amino-2-cyclopentylacetic acid (1 g, 6.98 mmol) in 1 M H₂SO₄(14 mL, 0.5 M) at 0° C. was added 2 M NaNO_(2(aq)) (11.5 mL, 10.5 mmol)slowly and the reaction was warmed to rt. After 48 hours, the reactionwas cooled back to 0° C., more 2 M NaNO₂ (10 mL, 9.11 mmol), and thereaction was warmed to rt. After 3 days, the reaction was extracted withEtOAc (3×50 mL), dried over Na₂SO₄ and concentrated in vacuo to afford63a (925 mg, 83%) as a white solid. ¹H NMR (400 MHz, d₆-DMSO): δ3.79 (d,J=5.9 Hz, 1H), 2.18-2.04 (m, 1H), 1.63-1.33 (m, 8H).

Compound 63b

To 1d (4 g, 9.24 mmol) in dioxane (5 mL) was added an HCl solution (34mL, 0.25 M, 4 M in dioxane). After stirring for 18 hours, the reactionwas concentrated in vacuo to afford the free dipeptide (3.69 g) as anamorphous, pale yellow solid. To 63a (501.5 mg, 3.48 mmol), dipeptide(1.28 g, 3.48 mmol), and i-Pr₂NEt (0.61 mL, 3.48 mmol) in DCM (35 mL,0.05 M) at 0° C. was added(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(1.25 g, 4.17 mmol) and the reaction was warmed to room temperature.After 24 hours, the reaction was concentrated in vacuo. The residue wasdiluted with EtOAc (200 mL), washed with 10% citric acid (2×100 mL),sat. NaHCO₃ (2×100 mL), and brine (200 mL), dried over Na₂SO₄, andconcentrated in vacuo. Purification by column chromatography equippedwith an ELSD (0-100% EtOAc/hexane) afforded 63b (458.3 mg, 20%) as agolden orange oil. ¹H NMR (400 MHz, CDCl₃): δ6.95 (s, 1H), 5.35 (p,J=6.8 Hz, 1H), 4.94 (d, J=12.0 Hz, 1H), 4.69 (d, J=12.0 Hz, 1H), 4.30(d, J=10.2 Hz, 1H), 4.07 (d, J=4.7 Hz, 1H), 3.69 (d, J=7.0 Hz, 1H), 2.92(m, 1H), 2.20 (m, 1H), 1.91 (m, 1H), 1.72 (m, 3H), 1.55 (m, 8H), 1.32(d, J=6.8 Hz, 3H). LCMS (m/z) 458.20 [M+H].

Compound 63c

To 39a (636 mg, 1.81 mmol) and 48f (286 mg, 1.81 mmol) in dioxane (14.5mL, 0.125 M) was added palladium (II) acetate (61.5 mg, 0.272 mmol),tris(2-methylphenyl)phosphine (168 mg, 0.543 mmol), triethylamine (0.71mL, 5.43 mmol). The mixture was heated in a microwave reactor to 100° C.for 60 minutes. The reaction was filtered through celite, rinsed withethyl acetate, and concentrated in vacuo. The crude material wassuspended in Et₂O (15 mL) and extracted with sat. NaHCO_(3(aq)) (9×30mL). The combined aqueous were acidified to ˜pH 4 using 1 M HCl_((aq)),extracted with EtOAc (3×500 mL), dried over MgSO₄, and concentrated invacuo to afford the title compound (457 mg, 59%) as a yellow foam. ¹HNMR (400 MHz, CDCl₃) δ 8.16 (m, 2H), 7.78 (s, 1H), 7.68 (s, 1H), 7.42(s, 1H), 6.90 (d, J=16.6 Hz, 1H), 6.38 (m, 2H), 5.01 (d, J=5.7 Hz, 2H),4.85 (d, J=6.1 Hz, 1H), 4.58 (m, 2H), 3.95 (m, 2H), 1.58 (s, 3H), 1.43(s, 9H). LCMS (m/z) 429.00 [M+H].

Compound 63d

To N-Boc protected 1,3-dioxane carboxylic acid 63c (149.7 mg, 0.35mmol), compound 63b (189.3 mg, 0.42 mmol), and DMAP (104.5 mg, 0.84mmol) in DCM (3.5 mL, 0.1 M) under Ar was added i-Pr₂NEt (0.15 mL, 0.84mmol) followed by 2-methyl-6-nitrobenzoic anhydride (181.6 mg, 0.525mmol). After 18 hours, the reaction was diluted with DCM (10 mL), washedwith water (3×10 mL), dried over MgSO₄, and concentrated in vacuo to ayellow foam that was purified via column chromatography (0-100%EtOAc/hexane) to afford 63d (192.0 mg, 51%) as an off-white foam. ¹H NMR(400 MHz, CDCl₃): δ8.04 (s, 1H), 7.96 (s, 1H), 7.71 (s, 1H), 7.54 (s,1H), 7.29 (d, J=7.4 Hz, 2H), 6.74 (d, J=16.7 Hz, 1H), 6.17 (d, J=16.2Hz, 2H), 5.35 (d, J=4.3 Hz, 1H), 5.32 (d, J=7.1 Hz, 1H), 5.16 (d, J=5.9Hz, 1H), 4.97 (d, J=6.1 Hz, 1H), 4.92 (d, J=12.0 Hz, 1H), 4.75 (d, J=5.9Hz, 1H), 4.68 (m, 3H), 4.31 (d, J=12.6 Hz, 1H), 3.73 (m, 5H), 2.85 (m,2H), 2.13 (m, 1H), 1.76 (m, 4H), 1.58 (m, 12H), 1.45 (s, 9H), 1.27 (d,J=6.9 Hz, 3H). LCMS (m/z) 868.15 [M+H].

Compound 63

To 63c (192.0 mg, 0.219 mmol) in THF (2.2 mL, 0.1 M) at 0° C. was added0.3 M NaOH_((aq)) (0.55 mL, 0.6.56 mmol). The reaction was monitored byLCMS hourly, with additions of 0.3 M NaOH_((aq)). The reaction was shownto be complete after 3 hours and was quenched with 1 M HCl_((aq)) (0.34mL) to ˜pH 4. The mixture was then diluted with EtOAc (15 mL), washedwith water (3×5 mL), dried over MgSO₄, and concentrated in vacuo toafford a yellow foam (171.4 mg). The crude material was dissolved indioxane (1 mL) and an HCl solution (3 mL, 4 M in dioxane) was added.After 60 min, the suspension was concentrated in vacuo to a yellow solid(193.1 mg). The crude material in DMF (1 mL) was diluted with DCM (100mL) and i-Pr₂NEt (0.22 mL, 1.23 mmol) was added, followed by HATU (113.8mg, 0.295 mmol). After 30 minutes, the mixture was washed with 5% LiCl(3×100 mL), sat. NaHCO₃ (100 mL), and brine (100 mL), dried over MgSO₄,and concentrated in vacuo to an orange residue. The crude material waspurified by column chrmoatography (25-100% EtOAc/hexane) to afford 63(46.0 mg, 26%) as a whitish residue. ¹H NMR (400 MHz, CDCl₃): δ9.04 (d,J=5.7 Hz, 1H), 8.16-8.03 (m, 4H), 7.72 (d, J=8.4 Hz, 2H), 7.48 (d, J=8.3Hz, 2H), 7.24 (s, 1H), 6.76 (d, J=16.4 Hz, 2H), 6.48 (d, J=8.5 Hz, 1H),6.14 (d, J=16.5 Hz, 1H), 5.90-5.79 (m, 1H), 5.15 (m, 2H), 5.00 (d, J=5.8Hz, 1H), 4.81 (d, J=6.0 Hz, 1H), 4.57 (m, 4H), 3.91 (dd, J=44.8, 11.5Hz, 3H), 3.56 (m, 2H), 2.57 (m, 2H), 2.43 (m, 2H), 1.62 (m, 8H). LCMS(m/z) 620.48 [M+H]. Rt=2.60 min.

Example 64 Compound 64 Example 64

Compound 64a

To 2-amino-2-cyclobutylacetic acid (1 g, 6.04 mmol) in 1 M H₂SO_(4(aq))(12 mL, 0.5 M) at 0° C. was added 2 M NaNO_(2(aq)) (15 mL, 30.19 mmol)slowly and the reaction was allowed to warm to rt slowly. After 3 days,¹H NMR showed no starting material and the reaction was extracted withEtOAc (3×50 mL). The combined organics were dried over Na₂SO₄ andconcentrated in vacuo to afford 64a (495.0 g, 44%) as a yellow solid. ¹HNMR (400 MHz, d₆-DMSO): δ3.78 (d, J=8.5 Hz, 1H), 2.62 (m, 1H), 1.94 (m,4H).

Compound 64b

To N-Boc protected quinolinyl 1,3-dioxane carboxylic acid 63c (105.3 mg,0.245 mmol), and a tripeptide (prepared using 64a from the methoddescribed for 1e) (129.9 mg, 0.294 mmol), and DMAP (73.5 mg, 0.588 mmol)in DCM (3 mL, 0.1 M) under argon was added i-Pr₂NEt (0.10 mL, 0.588mmol) followed by 2-methyl-6-nitrobenzoic anhydride (127.0 mg, 0.268mmol). After 18 hours, the reaction was diluted with DCM (10 mL), washedwith water (3×5 mL), dried over MgSO₄, and concentrated in vacuo to abrown foam that was purified via column chromatography (0-100%EtOAc/hexane) to afford 64b (201.3 mg, 85%) as an off-white foam. ¹H NMR(400 MHz, CDCl₃): δ8.06 (s, 1H), 7.98 (s, 1H), 7.73 (s, 1H), 7.54 (s,1H), 7.31 (s, 1H), 6.76 (m, 1H), 6.18 (m, 1H), 5.32 (m, 2H), 5.17 (dd,J=17.5, 5.9 Hz, 1H), 4.90 (m, 2H), 4.74 (m, 4H), 4.38 (m, 1H), 3.79 (m,5H), 3.00 (m, 1H), 2.86 (d, J=10.3 Hz, 1H), 2.14 (m, 2H), 1.88 (m, 8H),1.62 (m, 4H), 1.53 (s, 3H), 1.46 (s, 9H). LCMS (m/z) 856.16 [M+H].

Compound 64

To 64b (201.3 mg, 0.235 mmol) in THF (2.5 mL, 0.1 M) at 0° C. was added0.3 M NaOH_((aq)) (0.58 mL, 0.177 mmol). The reaction was monitored byLCMS hourly, with additions of 0.3 M NaOH_((aq)). The reaction was shownto be complete after 90 minutes and was quenched with 1 M HCl_((aq))(0.26 mL) to ˜pH 4. The mixture was then diluted with EtOAc (15 mL),washed with water (3×5 mL), dried over MgSO₄, and concentrated in vacuoto afford a white foam. The crude material was dissolved in dioxane (1mL) and an HCl solution (3 mL, 4 M in dioxane) was added. After 60 min,the suspension was concentrated in vacuo to a yellow solid. The crudematerial in DMF (0.5 mL) was diluted with DCM (100 mL) and i-Pr₂NEt(0.22 mL, 1.26 mmol) was added followed, by HATU (119.6 mg, 0.302 mmol).After 30 minutes, the mixture was washed with 5% LiCl_((aq)) (3×100 mL),sat. NaHCO₃ (100 mL), and brine (100 mL), dried over MgSO₄, andconcentrated in vacuo. The crude material was purified by prep HPLC(Synergi Polar-RP, 40-55% MeCN/H₂O) to afford 64 (3.5 mg, 2%) as awhitish residue. ¹H NMR (400 MHz, CDCl₃): δ9.48 (s, 1H), 8.60 (s, 1H),8.52 (s, 1H), 7.96 (s, 2H), 7.66 (d, J=8.6 Hz, 1H), 6.85 (d, J=16.7 Hz,1H), 6.35 (d, J=16.6 Hz, 1H), 6.20 (d, J=8.6 Hz, 1H), 5.53 (m, 1H), 5.30(m, 1H), 4.94 (dd, J=7.2, 3.2 Hz, 2H), 4.87 (d, J=5.9 Hz, 1H), 4.48 (m,J=9.8 Hz, 3H), 4.06 (d, J=11.2 Hz, 1H), 3.94 (d, J=11.1 Hz, 1H), 3.76(m, 2H), 2.68 (m, 1H), 2.54 (m, 1H), 1.92 (m, 13H), 1.64 (m, 6H). LCMS(m/z) 606.46 [M+H]. Tr=3.57 min (Gemini 5 u C18 110 Å, 50×4.60 mm 5micron column, 10 min, 2 ml/min, 5-100% acetonitrile/water, 0.1% TFAmodifier).

Example 65 Compound 65

Compound 65a

To L-isoleucine (2 g, 15.2 mmol) in 1 M H₂SO_(4(aq)) (30 mL, 0.5 M) at0° C. was added 2 M NaNO_(2(aq)) (11.5 mL, 22.9 mmol) slowly and thereaction was allowed to warm to rt slowly. After 7 days, the reactionwas saturated with (NH₄)₂SO₄, extracted with EtOAc (3×50 mL), dried overNa₂SO₄ and concentrated in vacuo to afford 65a (1.25 g, 57%) as a whitesolid. ¹H NMR (400 MHz, d₆-DMSO): δ3.77 (d, J=4.9 Hz, 1H), 1.66 (m, 1H),1.40 (m, 1H), 1.15 (1H), 0.84 (dd, J=15.0, 7.2 Hz, 6H).

Compound 65b

To 1d (5.88 g, 13.6 mmol) was added an HCl solution (13.6 mL, 1 M, 4 Min dioxane). After stirring for 18 hours, the reaction was concentratedin vacuo to afford the free dipeptide as an amorphous, pale yellowsolid. To 65a (492.4 mg, 3.78 mmol), dipeptide (1.39 g, 3.78 mmol), andi-Pr₂NEt (0.66 mL, 3.787 mmol) in DCM (40 mL, 0.1 M) at 0° C. was added(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(1.38 g, 4.54 mmol) and the reaction was slowly warmed to roomtemperature. After 18 hours, the reaction was concentrated in vacuo. Theresidue was diluted with EtOAc (200 mL), washed with 10% citric acid(200 mL), sat. NaHCO₃ (200 mL), and brine (200 mL), dried over Na₂SO₄,and concentrated in vacuo. Purification by column chromatographyequipped with an ELSD (0-100% EtOAc/hexane) afforded 65b (856.8 mg, 51%)as a white solid. ¹H NMR (400 MHz, CDCl₃): δ7.18 (s, 1H), 5.39 (m, 1H),4.95 (d, J=11.9 Hz, 1H), 4.71 (d, J=11.9 Hz, 1H), 4.34 (m, 1H), 4.00 (d,J=3.4 Hz, 1H), 3.70 (m, 1H), 2.93 (m, 1H), 2.18 (m 1H), 1.89 (m, 1H),1.73 (t, J=9.7 Hz, 2H), 1.42 (m, 1H), 1.34 (d, J=6.9 Hz, 3H), 1.24 (t,J=8.7, 5.6 Hz, 3H), 1.19 (m, 1H), 1.00 (d, J=6.9 Hz, 3H). LCMS (m/z)46.00 [M+H].

Compound 65c

To the N-Boc protected quinolinyl 1,3-dioxane carboxylic acid 63c (74.9mg, 0.175 mmol), 65b (96.2 mg, 0.175 mmol), and DMAP (52.7 mg, 0.420mmol) in DCM (1.8 mL, 0.1 M) under Ar was added i-Pr₂NEt (0.07 mL, 0.420mmol) followed by 2-methyl-6-nitrobenzoic anhydride (93.1 mg, 0.263mmol). After 18 hours, the reaction was diluted with DCM (10 mL), washedwith water (3×5 mL), dried over MgSO₄, and concentrated in vacuo toafford 65c as a yellow foam (185.8 mg, 99%). ¹H NMR (400 MHz, CDCl₃):δ8.22 (s, 1H), 8.06 (d, J=8.4 Hz, 1H), 7.98 (s, 1H), 7.73 (d, J=8.4 Hz,1H), 7.31 (d, J=8.4 Hz, 1H), 6.75 (d, J=16.7 Hz, 1H), 6.55 (s, 2H), 6.18(d, J=16.2 Hz, 1H), 5.34 (m, 2H), 4.98 (m, 2H), 4.73 (m, 4H), 4.33 (d,J=14.7 Hz, 1H), 3.85 (dd, J=27.7, 11.0 Hz, 2H), 2.18 (dt, J=15.3, 8.4Hz, 1H), 1.53 (d, J=6.6 Hz, 3H), 1.47 (s, 9H), 1.33 (d, J=6.8 Hz, 2H),1.29 (d, J=6.8 Hz, 3H), 0.98 (d, J=6.8 Hz, 3H), 0.92 (t, J=7.6 Hz, 3H).LCMS (m/z) 856.51 [M+H].

Compound 65

To 65c (150 mg, 0.175 mmol) in THF (1.8 mL, 0.1 M) at 0° C. was added0.3 M NaOH_((aq)) (0.44 mL, 0.131 mmol). The reaction was monitored byLCMS ever 30 minutes, with additions of 0.3 M NaOH_((aq)). The reactionwas shown to be complete after 2 hours and was quenched with 1 MHCl_((aq)) (0.40 mL) to ˜pH 4. The mixture was then diluted with EtOAc(15 mL), washed with water (3×5 mL) and brine (5 mL), dried over MgSO₄,and concentrated in vacuo to afford a yellow foam (116.8 mg). The crudematerial was dissolved in dioxane (1 mL) and an HCl solution (3 mL, 4 Min dioxane) was added. After 60 min, the suspension was concentrated invacuo to an orange solid. The crude material in DMF (1 mL) was dilutedwith DCM (100 mL) and i-Pr₂NEt (0.22 mL, 1.26 mmol) was added, followedby HATU (305.2 mg, 0.803 mmol). After 30 minutes, the mixture was washedwith 5% LiCl_((aq)) (3×100 mL), sat. NaHCO_(3(aq)) (100 mL), and brine(100 mL), dried over MgSO₄, and concentrated in vacuo to an orangeresidue. The crude material was purified by column chromatography(20-100% MeCN/H₂O). The residue was dissolved in EtOAc (20 mL) andwashed with 5% LiCl (10×10 mL) to afford 65 (24.2 mg, 13%) as a whitishresidue. ¹H NMR (400 MHz, CDCl₃): δ9.09 (s, 1H), 8.11 (s, 1H), 7.75 (s,1H), 7.52 (s, 1H), 7.3 (s, 1H), 6.73 (d, J=16.5 Hz, 2H), 6.33 (d, J=8.6Hz, 1H), 6.14 (d, J=16.2 Hz, 1H), 5.85 (s, 1H), 5.11 (s, 2H), 5.02 (d,J=5.9 Hz, 1H), 4.80 (d, J=6.2 Hz, 1H), 4.65 (d, J=11.3 Hz, 1H), 4.56 (d,J=11.6 Hz, 2H), 3.95 (d, J=11.5 Hz, 1H), 3.83 (d, J=11.3 Hz, 1H), 3.56(m, 1H), 2.56 (m, 1H), 1.96 (m, 1H), 1.61 (m, 10H), 1.26 (t, J=7.1 Hz,3H), 1.01 (d, J=5.8 Hz, 3H). LCMS (m/z) 608.47 [M+H]. Tr=3.67 min.

Example 66 Compound 66

Compound 66a

To 1d (4 g, 9.24 mmol) in dioxane (5 mL) was added an HCl solution (34mL, 0.25 M, 4 M in dioxane). After stirring for 18 hours, the reactionwas concentrated in vacuo to afford the free dipeptide (3.69 g) as anamorphous, pale yellow solid. To α-hydroxy cyclopropyl acetic acid(obtained from ArkPharm, Inc.), (493.5 mg, 4.09 mmol), dipeptide (1000mg, 2.72 mmol), and i-Pr₂NEt (0.48 mL, 2.72 mmol) in DCM (45 mL, 0.05 M)at 0° C. was added (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (981.1 mg, 3.27 mmol) and the reaction was slowlywarmed to room temperature. After 24 hours, the reaction wasconcentrated in vacuo. The residue was diluted with EtOAc (100 mL),washed with 10% citric acid (5×50 mL), sat. NaHCO_(3(aq)) (2×50 mL), andbrine (100 mL), dried over Na₂SO₄, and concentrated in vacuo.Purification by column chromatography equipped with an ELSD (0-100%EtOAc/hexane) afforded 66a (425.8 mg, 36%) as a white foam. ¹H NMR (400MHz, CDCl₃): δ7.14 (m, 1H), 5.37 (m, 1H), 4.95 (d, J=12.0 Hz, 1H), 4.72(dd, J=12.0, 2.8 Hz, 1H), 4.33 (m, 1H), 3.86 (m, 1H), 3.70 (m, 1H), 3.48(dd, J=12.0, 8.2 Hz, 1H), 2.93 (m, 1H), 2.86 (d, J=10.3 Hz, 1H), 2.19(m, 1H), 1.94 (m, 1H), 1.73 (m, 2H), 1.35 (m, 3H), 1.09 (m, 1H), 0.67(m, 1H), 0.54 (m, 3H). LCMS (m/z) 430.12 [M+H].

Compound 66b

To the indicated N-Boc quinolinyl carboxylate 74c (242.1 mg, 0.451mmol), 66a (238.4 mg, 0.451 mmol), and DMAP (132.3 mg, 1.08 mmol) in DCM(4.6 mL, 0.1 M) under Ar was added i-Pr₂NEt (0.19 mL, 1.08 mmol)followed by 2-methyl-6-nitrobenzoic anhydride (235.6 mg, 0.676 mmol).After 18 hours, the reaction was diluted with DCM (10 mL), washed withwater (3×10 mL), dried over MgSO₄, and concentrated in vacuo to a yellowfoam that was purified via column chromatography (0-100% EtOAc/hexane)to afford 66b (313.7 mg, 66%) as a pale golden oil. ¹H NMR (400 MHz,CDCl₃): δ 8.05 (d, J=8.3 Hz, 1H), 7.97 (s, 1H), 7.70 (d, J=8.3 Hz, 1H),7.62 (d, J=8.2 Hz, 1H), 7.27 (s, 1H), 7.08 (m, 1H), 6.76 (d, J=16.6 Hz,1H), 6.43 (d, J=16.3 Hz, 1H), 6.19 (m, 1H), 5.25 (m, J=15.1 Hz, 1H),4.92 (m, 2H), 4.81 (dd, J=14.1, 6.4 Hz, 1H), 4.70 (dd, J=12.7, 10.3 Hz,1H), 4.29 (m, 1H), 3.66 (m, 3H), 2.49 (m, 1H), 2.09 (m, 2H), 1.80 (m,2H), 1.68 (m, 4H), 1.52 (d, J=6.2 Hz, 3H), 1.47 (s, 9H), 1.29 (m, 3H),1.12 (s, 1H), 0.57 (m, 4H). LCMS (m/z) 968.24 [M+H].

Compound 66

To 66b (313.7 mg, 0.324 mmol) in THF (3.2 mL, 0.1 M) at 0° C. was added0.3 M NaOH (0.81 mL, 0.243 mmol). The reaction was monitored by LCMSever 30 minutes, with additions of 0.3 M NaOH. The reaction was shown tobe complete after 60 minutes and was quenched with 1 M HCl (0.36 mL) to˜pH 4. The mixture was then diluted with EtOAc (15 mL), washed withwater (3×5 mL), dried over MgSO₄, and concentrated in vacuo to afford apale yellow foam (283.7 mg). The crude material was dissolved in dioxane(1 mL) and an HCl solution (3 mL, 4 M in dioxane) was added. After 60min, the suspension was concentrated in vacuo to a pale yellow solid(275.5 mg). The crude material in DMF (1.5 mL) was diluted with DCM (100mL) and i-Pr₂NEt (0.30 mL, 1.69 mmol) was added, followed by HATU (157.2mg, 0.405 mmol). After 30 minutes, the mixture was washed with 5%LiCl_((aq)) (3×100 mL), sat. NaHCO₃ (100 mL), and brine (100 mL), driedover MgSO₄, and concentrated in vacuo. The crude material was purifiedby prep HPLC (Synergi Polar-RP, 35% MeCN/H₂O) and lyophilized to afford66 (3.4 mg, 1%) as a white solid. ¹H NMR (400 MHz, CDCl₃): δ8.54 (s,1H), 8.10 (s, 1H), 7.73 (d, J=7.3 Hz, 2H), 7.43 (s, 1H), 6.72 (dd,J=19.3, 8.7 Hz, 1H), 6.48 (dd, J=24.9, 16.4 Hz, 1H), 5.89 (m, 1H), 5.17(t, J=6.2 Hz, 1H), 4.72 (m, 1H), 4.57 (m, 1H), 3.70 (m, 2H), 3.52 (m,2H), 2.54 (m, 6H), 2.02 (m, 3H), 1.62 (m, 10H), 1.30 (m, 3H), 0.88 (m,1H), 0.60 (m, 4H). LCMS (m/z) 604.60 [M+H]. Tr=2.23 min (Gemini 5 u C18110 Å, 50×4.60 mm 5 micron column, 5 min, 2 ml/min, 5-100%acetonitrile/water, 0.1% TFA modifier).

Example 67 Compound 67

Compound 67a

To (S)-2-amino-2-(tetrahydro-2H-pyran-4-yl)acetic acid (obtained fromAstaTech, Inc.), (1.01 g, 6.35 mmol) in 1 M H₂SO_(4(aq)) (12.5 mL, 0.5M) at 0° C. was added 2 M NaNO_(2(aq)) (9.5 mL, 19.0 mmol) slowly andthe reaction was allowed to warm to rt slowly. After 24 hours, ¹H NMRshowed no starting material and the reaction was extracted with EtOAc(3×50 mL). The combined organics were dried over Na₂SO₄ and concentratedin vacuo to afford 67a (868.8 g, 85%) as an off-white solid. ¹H NMR (400MHz, d₆-DMSO): δ3.84 (m, 2H), 3.76 (d, J=5.1 Hz, 1H), 3.24 (m, 2H), 1.81(m, 1H), 1.39 (m, 4H).

Compound 67b

To 1d (5 g, 11.6 mmol) in dioxane (5 mL) was added an HCl solution (45mL, 0.25 M, 4 M in dioxane). After stirring for 18 hours, the reactionwas concentrated in vacuo to afford the free dipeptide (4.71 g) as anamorphous, pale yellow solid.

To 67a (300.9 mg, 1.87 mmol), dipeptide (447.9 mg, 1.25 mmol), andi-Pr₂NEt (0.27 mL, 1.56 mmol) in DCM (16 mL, 0.1 M) at 0° C. was added(benzotriazol-1-(benzyloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (827.9 mg, 1.87 mmol) and the reaction was slowlywarmed to room temperature. After 18 hours, the reaction wasconcentrated in vacuo. The residue was diluted with EtOAc (100 mL),washed with 10% citric acid (3×100 mL), sat. NaHCO_(3(aq)) (100 mL), andbrine (100 mL), dried over Na₂SO₄, and concentrated in vacuo.Purification by column chromatography equipped with an ELSD (0-100%EtOAc/hexane) afforded 67b (171.3 mg, 23%) as an off-white foam. ¹H NMR(400 MHz, CDCl₃): δ7.15 (d, J=7.9 Hz, 1H), 5.38 (p, J=6.8 Hz, 1H), 4.95(d, J=11.9 Hz, 1H), 4.71 (d, J=11.9 Hz, 1H), 4.31 (d, J=14.2 Hz, 1H),3.99 (m, 3H), 3.94 (d, J=3.8 Hz, 1H), 3.70 (m, 1H), 3.39 (m, 2H), 2.94(m, 1H), 2.19 (m, 1H), 1.98 (m, 2H), 1.75 (dd, J=19.3, 10.0 Hz, 2H),1.64 (ddd, J=24.9, 12.7, 4.6 Hz, 2H), 1.55 (t, J=8.4 Hz, 2H), 1.39 (m,1H), 1.33 (d, J=6.8 Hz, 3H). LCMS (m/z) 474.26 [M+H].

Compound 67c

To (R,E)-4-(2-(1-(tert-butoxycarbonylamino)ethyl)quinolin-7-yl)-2,2dimethylbut-3-enoic acid (136.9 mg, 0.361 mmol), 67b (171.3 mg, 0.361mmol), and DMAP (106.1 mg, 866 mmol) in DCM (3.5 mL, 0.1 M) under Ar wasadded i-Pr₂NEt (0.15 mL, 866 mmol) followed by 2-methyl-6-nitrobenzoicanhydride (189.2 mg, 0.541 mmol). After 2.5 hours, more2-methyl-6-nitrobenzoic anhydride (50.8) was added. After another hour,the reaction was diluted with DCM (10 mL), washed with water (3×5 mL),dried over MgSO₄, and concentrated in vacuo to afford 67c as anoff-white foam (324.4 mg, 90%). ¹H NMR (400 MHz, CDCl₃): δ8.22 (d, J=5.9Hz, 2H), 8.05 (d, J=8.3 Hz, 1H), 8.02 (s, 1H), 7.71 (s, 1H), 7.66 (d,J=8.8 Hz, 1H), 7.28 (s, 1H), 7.12 (s, 1H), 6.70 (d, J=14.1 Hz, 1H), 6.52(d, J=6.4 Hz, 2H), 6.22 (s, 1H), 5.14 (d, J=3.7 Hz, 1H), 4.94 (m, 2H),4.70 (d, J=12.1 Hz, 1H), 3.96 (s, 2H), 3.73 (m, 1H), 3.37 (t, J=12.1 Hz,2H), 2.29 (m, 1H), 2.19 (m, 1H), 1.89 (m, 1H), 1.57 (d, J=6.1 Hz, 6H),1.52 (m, 6H), 1.48 (m, 15H), 1.28 (d, J=6.7 Hz, 3H). LCMS (m/z) 840.67[M+H].

Compound 67

To 67c (273.5 mg, 0.325 mmol) in THF (3.4 mL, 0.1 M) at 0° C. was added0.3 M aq. NaOH (8.2 mL, 0.244 mmol). The reaction was monitored by LCMShourly, with additions of 0.3 M aq. NaOH. The reaction was shown to becomplete after 2 hours and was quenched with 1 M aq. HCl (0.77 mL) to˜pH 4. The mixture was then diluted with EtOAc (15 mL), washed withwater (3×5 mL) and brine (5 mL), dried over MgSO₄, and concentrated invacuo to afford a yellow foam (251.0 mg). The crude material wasdissolved in dioxane (1 mL) and HCl (3 mL, 4 M in dioxane) was added.After 60 min, the suspension was concentrated in vacuo to a yellow solid(266.8 mg). The crude material in DMF (1 mL) was diluted with DCM (100mL) and i-Pr₂NEt (0.28 mL, 1.63 mmol) was added, followed by HATU (148.9g, 0.325 mmol). After 30 minutes, the mixture was washed with 5% aq.LiCl (3×100 mL), sat. NaHCO_(3(aq)) (100 mL), and brine (100 mL), driedover MgSO₄, and concentrated in vacuo to an orange residue. The crudematerial was purified by column chromatography (25-100% EtOAc/hexane) toafford 67 (38.4 mg, 18%) as a pale yellow residue. ¹H NMR (400 MHz,CDCl₃): δ9.07 (s, 1H), 8.13 (d, J=7.1 Hz, 1H), 8.05 (s, 1H), 7.74 (d,J=8.1 Hz, 1H), 7.47 (d, J=7.4 Hz, 1H), 7.28 (s, 1H), 6.65 (d, J=16.0 Hz,1H), 6.43-6.26 (m, 1H), 5.79 (m, 1H), 5.36 (m, 1H), 5.23 (m, 1H), 5.11(m, 1H), 4.99 (d, J=7.6 Hz, 1H), 4.54 (dd, J=13.5, 2.5 Hz, 1H), 3.96 (d,J=11.1 Hz, 2H), 3.84 (dd, J=33.2, 14.1 Hz, 1H), 3.64 (d, J=11.9 Hz, 1H),3.51 (t, J=11.5 Hz, 1H), 3.33 (m, 3H), 2.57 (td, J=13.1, 2.5 Hz, 1H),2.43 (d, J=14.1 Hz, 1H), 2.33 (m, 1H), 2.21 (m, 1H), 1.96 (d, J=12.9 Hz,1H), 1.70 (m, 4H), 1.63 (s, 6H), 1.60 (m, 6H), 1.53 (s, 3H), 1.48 (s,2H), 1.44 (s, 3H). LCMS (m/z) 592.37 [M+H]. Tr=2.45 min.

Example 68 Compound 68

Compound 68a

To 2-amino-3-fluorobutanoic acid (obtained from Fluorochem, Ltd.),(509.4 mg, 4.13 mmol) in 1 M H₂SO_(4(aq)) (8.5 mL, 0.5 M) at 0° C. wasadded 2 M NaNO_(2(aq)) (6.0 mL, 12.4 mmol) slowly and the reaction wasallowed to warm to rt slowly. After 24 hours, ¹H NMR showed tracestarting material and the reaction was extracted with EtOAc (3×25 mL).The combined organics were dried over Na₂SO₄ and concentrated in vacuoto afford 68a (437.2 g, 87%) as a white solid. ¹H NMR (400 MHz,d₆-DMSO): δ5.07-4.65 (m, 1H), 4.07 (ddd, J=33.7, 23.1, 3.1 Hz, 1H), 1.28(ddd, J=24.7, 18.4, 6.4 Hz, 3H). ¹⁹F NMR (376 MHz, DMSO): δ−180.63 (dqd,J=48.5, 24.3, 14.9 Hz), −186.54 (ddq, J=48.3, 31.5, 24.1 Hz).

Compound 68b

To 1d (5 g, 11.6 mmol) in dioxane (5 mL) was added an HCl solution (45mL, 0.25 M, 4 M in dioxane). After stirring for 18 hours, the reactionwas concentrated in vacuo to afford the free dipeptide (4.71 g) as anamorphous, pale yellow solid. To 68a (225.3 mg, 1.84 mmol), dipeptide(671.2 mg, 1.84 mmol), and i-Pr₂NEt (0.32 mL, 1.84 mmol) in DCM (20 mL,0.1 M) at 0° C. was added(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(1.2130 g, 2.76 mmol) and the reaction was slowly warmed to roomtemperature. After 18 hours, the reaction was concentrated in vacuo. Theresidue was diluted with EtOAc (100 mL), washed with 10% citric acid(3×100 mL), sat. NaHCO_(3(aq)) (100 mL), and brine (100 mL), dried overNa₂SO₄, and concentrated in vacuo. Purification by column chromatographyequipped with an ELSD (0-100% EtOAc/hexane) afforded 68b (318.9 mg, 32%)as a white foam. ¹H NMR (400 MHz, CDCl₃): δ5.39 (m, 1H), 5.07 (m 1H),4.95 (d, J=11.9 Hz, 1H), 4.70 (dd, J=11.9, 4.2 Hz, 1H), 4.31 (m, 1H),4.10 (ddd, J=13.9, 10.0, 5.5 Hz, 1H), 3.70 (d, J=6.4 Hz, 1H), 2.91 (m,1H), 2.87 (d, J=10.3 Hz, 1H), 2.20 (m, 1H), 1.91 (m, 1H), 1.72 (m, 1H),1.36 (m, 8H). ¹⁹F NMR (376 MHz, CDCl₃): δ−181.98-−182.68 (m),−186.38-−187.09 (m), −187.09-−187.99 (m). LCMS (m/z) 437.82 [M+H].

Compound 68c

To the indicated N-Boc-quinolinyl carboxylate (211.3 mg, 0.538 mmol),68b (235.0 mg, 0.538 mmol), and DMAP (156.9 mg, 1.29 mmol) in DCM (5.4mL, 0.1 M) under Ar was added i-Pr₂NEt (0.23 mL, 1.29 mmol) followed by2-methyl-6-nitrobenzoic anhydride (279.5 mg, 0.807 mmol). After 18hours, the reaction was diluted with DCM (15 mL), washed with water (3×5mL), dried over MgSO₄, and concentrated in vacuo to afford 68c as a paleyellow foam (455.7 mg, 90%). ¹H NMR (400 MHz, CDCl₃): δ 8.10 (d, J=8.7Hz, 1H), 8.03 (m, 1H), 7.68 (d, J=21.0 Hz, 2H), 7.27 (s, 1H), 6.74 (d,J=7.5 Hz, 2H), 6.21 (m, 1H), 5.27 (d, J=15.8 Hz, 2H), 5.12 (m, 1H), 4.93(d, J=12.0 Hz, 1H), 4.70 (dd, J=11.9, 2.9 Hz, 1H), 4.28 (m, 1H), 3.69(m, 1H), 2.86 (d, J=10.2 Hz, 1H), 2.65 (d, J=9.3 Hz, 1H), 2.16 (M, 1H),1.89 (M, 1H), 1.63 (s, 2H), 1.61 (d, J=3.7 Hz, 3H), 1.54 (d, J=10.1 Hz,9H), 1.47 (s, 10H), 1.29 (m, 8H).

Compound 68

To 68c (455.7 mg, 0.567 mmol) in THF (5.4 mL, 0.1 M) at 0° C. was added0.3 M aq. NaOH (1.3 mL, 0.426 mmol). The reaction was monitored by LCMShourly, with additions of 0.3 M aq. NaOH. The reaction was shown to becomplete after 3 hours and was quenched with 1 M aq. HCl (0.75 mL) to˜pH 4. The mixture was then diluted with EtOAc (15 mL), washed withwater (3×5 mL) and brine (2×5 mL), dried over MgSO₄, and concentrated invacuo to afford a pale yellow foam (388.7 mg). The crude material wasdissolved in dioxane (1.5 mL) and an HCl solution (3 mL, 4 M in dioxane)was added. After 60 min, the suspension was concentrated in vacuo to ayellow solid (464.1 mg). The crude material in DMF (1.5 mL) was dilutedwith DCM (100 mL) and i-Pr₂NEt (0.51 mL, 2.90 mmol) was added, followedby HATU (265.6 mg, 0.695 mmol). After 30 minutes, the mixture was washedwith 5% LiCl_((aq)) (3×100 mL), sat. NaHCO_(3(aq)) (100 mL), and brine(100 mL), dried over MgSO₄, and concentrated in vacuo to an orangeresidue. The crude material was purified by prep HPLC (Synergi Polar-RP,50% MeCN/H₂O) and lyophilized to afford 68 (25.3 mg, 7%) as a whitesolid. ¹H NMR (400 MHz, CDCl₃): δ9.01 (d, J=9.8 Hz, 1H), 8.22 (s, 1H),8.10 (s, 1H), 7.74 (d, J=8.2 Hz, 1H), 7.46 (d, J=7.6 Hz, 1H), 6.97 (m,1H), 6.70 (m, 2H), 6.48 (d, J=15.7 Hz, 1H), 5.86 (m, 1H), 5.37 (m, 1H),5.18 (m, 1H), 4.55 (d, J=11.9 Hz, 1H), 3.83 (m, 1H), 3.57 (m, 1H), 3.27(m, 1H), 2.52 (m, 1H), 1.95 (m, 1H), 1.68 (d, J=22.6 Hz, 6H), 1.59 (s,6H), 1.55 (s, 2H), 1.48 (s, 6H), 1.43-1.21 (m, 3H). ¹⁹F NMR (376 MHz,CDCl₃) δ −184.47-−185.27 (m), −187.78-−188.41 (m). LCMS (m/z) 554.42[M+H]. Tr=2.49.

Example 69 Compound 69

Compound 69a

Beginning from 18e (192 mg, 0.50 mmol), N-Boc amine protection andsubsequent ester hydrolysis with NaOH/THF/water gave, afteracidification with 1N HCl and concentration to a crude residue, theintermediate carboxylic acid in quantitative yield. The resultingmaterial was carried forward directly into amide formation where2-methyl-6-nitrobenzoic anhydride (344 mg, 1 mmol),4-dimethylaminopyridine (128 mg, 1.05 mmol), and the N-Boc protectedisoquinolinyl acid were stirred in anhydrous dichloromethane (20 mL).Into the resulting solution was added N,N-diisopropylethylamine (0.26mL, 1.50 mmol) and this reaction mixture was stirred at room temperaturefor 10 minutes. Following the procedure described for Example 58, le(331 mg, 0.75 mmol) was added dropwise in anhydrous dichloromethane (10mL). After stirring for 12 hours at room temperature, the reactionmixture was transferred to a separatory funnel and washed with water (20mL, 10 mL of brine was added to support the separation). The aqueousphase was extracted with dichloromethane (20 mL). Combined organicextracts were washed with brine (20 mL) and dried over magnesiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by silica gel chromatography (gradient from 0-40% ethylacetate+methanol (4/1) in iso-hexanes) to afford 69a (391 mg, 97%) as awhite solid after evaporation. LCMS (m/z) 798.1 [M+H]′ Tr=2.82 min(Gemini 5 u C18 110 Å, 50×4.60 mm 5 micron column, 3.5 min, 2 ml/min,5-100% acetonitrile/water, 0.1% acetic acid modifier gradient).

Compound 69

To 69a (390 mg, 0.48 mmol) in tetrahydrofuran (20 mL) was added asolution of lithium hydroxide (13 mg, 0.53 mmol) in water (10 mL). Afterstirring at room temperature for 2 hour, 1M hydrochloric acid was added(0.55 mL of 1M solution in water, 0.55 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (10 mL, 40 mmol) at room temperature under argon for 4hours. Reaction mixture was concentrated under reduced pressure. Thisresidue was dissolved in N,N-dimethylformamide (5 mL) and the obtainedsolution was added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (276 mg, 0.73 mmol),N,N-diisopropylethylamine (312 mg, 2.42 mmol) and dichloromethane (200mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (100 ml) and brine (100 mL), dried over magnesium sulfate,filtered and concentrated under reduced pressure. The residue waspurified by silica gel chromatography (gradient from 0-40% ethylacetate+methanol (4/1) in iso-hexanes) to afford 69 (4 mg, 3%) as awhite solid after evaporation. ¹H NMR (400 MHz, CD₃CN): δ 9.12 (s, 1H),7.93 (d, J=6.4 Hz, 1H), 7.60, (s, 1H), 7.43 (s, 1H), 7.27 (m, 1H), 7.03(d, J=12 Hz, 1H), 6.83 (d, J=12, 1H), 5.65 (d, J=3 Hz, 1H), 4.96 (m,2H), 4.30 (d, J=9.3 Hz, 1H), 3.21 (m, 1H), 2.91 (bm, 1H), 2.32 (m, 1H),1.94-1.75 (cm, 3H), 1.64 (s, 3H), 1.62-1.48 (m, 4H), 1.46-1.35 (m, 2H),1.01 (d, J=6.7 Hz, 3H), 0.91 (d, J=6.7 Hz, 3H). LCMS (m/z) 550.4 [M+H]′Tr=1.81 min.

Example 70 Compound 70

Compound 70a

To a solution of 3,3-dimethyl butanoic acid (1 g, 7.5 mmol) in 1Msulfuric acid (15 ml, 1M aqueous solution), cooled to 0° C., was added asolution of sodium nitrite (1.0 g, 15 mmol) in water (8 ml). Thetemperature was maintained below 50° C. during the addition, and themixture was stirred at such overnight. The solution was then saturatedwith ammonium sulfate, extracted with diethyl ether (5×25 ml), driedover sodium sulfate and evaporated under reduced pressure giving thetitle compound (0.27 g, 67%) as a colorless oil that crystallized onstanding. ¹H NMR (400 MHz, CDC₃) 3.41 (s, 1H), 1.25 (s, 9H). LCMS (m/z)133.2 [M+H], Tr=0.39 min (Gemini 5 u C18 110 Å, 50×4.60 mm 5 microncolumn, 3.5 min, 2 ml/min, 5-100% acetonitrile/water, 0.1% acetic acidmodifier gradient).

Compound 70b

A solution of 1d (517 mg, 1.19 mmol) in dichloromethane (20 mL) wascooled in an ice water bath. Trimethylsilyl trifluoromethanesulfonate(398 mg, 1.79 mmol) was added dropwise at 0° C. under argon, and theresulting solution was stirred at room temperature for 30 minutes. Thereaction mixture was evaporated to dryness and the resulting cruderesidue (LCMS (m/z) 332.2/334.3 [M+H]′ Tr=2.06 min) was dissolved inanhydrous dichloromethane (20 mL) and cooled to 0° C. To this was added70a (192 mg, 1.43 mmol), N,N-diisopropylethylamine (0.51 mL, 2.98 mmol),and (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (633 mg, 1.43 mmol). The reaction was allowed towarm to room temperature and stirred for 8 hours. The mixture wasevaporated to dryness and the residue was dissolved in ethyl acetate(100 mL) and washed with 10% citric acid (100 mL), saturated NaHCO₃ (100mL), and brine (100 mL). The organic layer was dried over MgSO₄, onevolume equivalent of hexane was added and this solution was filteredthrough a 5 cm layer of silica gel, silica gel layer was washed with 50mL of ethyl acetate/hexane mixture (1/1). The desired product was washedout with ethyl acetate (100 mL), concentrated under reduced pressure andco-distilled with dichloromethane. The 70b compound (537 mg,quantitative yield) was isolated after drying under high vacuum for oneday. ¹H NMR (400 MHz, CD₃OD): δ 5.52 (q, J=6.9 Hz, 1H), 5.11 (d, J=12.1Hz, 1H), 4.91 (d, J=12.2 Hz, 1H), 3.94 (d, J=2.6 Hz, 1H), 3.84 (dd,J=7.1, 4.7 Hz, 1H), 3.84 (m, 1H), 3.80-3.58 (m, 2H), 3.41 (s, 3H),2.33-2.13 (m, 1H), 2.09-1.92 (m, 2H), 1.86-1.75 (m, 1H), 1.42 (d, J=6.9Hz, 3H), 1.32 (s, 9H). LCMS (m/z) 446.0[M+H]′ Tr=2.11 min.

Compound 70c

Into an oven-dried, argon purged flask were placed2-methyl-6-nitrobenzoic anhydride (172 mg, 0.5 mmol),4-dimethylaminopyridine (64 mg, 0.50 mmol), the indicatedN-Boc-quinolinyl carboxylate (96 mg, 0.25 mmol), and anhydrousdichloromethane (10 mL). Into the resulting solution was addedN,N-diisopropylethylamine (0.13 mL, 0.75 mmol) and this reaction mixturewas stirred at room temperature for 10 minutes. 70b (168 mg, 0.38 mmol)was added dropwise via syringe as a solution in anhydrousdichloromethane (10 mL). After stirring for 12 hours at roomtemperature, the reaction mixture was transferred to a separatory funneland washed with water (20 mL, 10 mL of brine was added to support theseparation). The aqueous phase was extracted with dichloromethane (20mL). Combined organic extracts were washed with brine (20 mL) and driedover magnesium sulfate, filtered and concentrated under reducedpressure. The residue was purified by silica gel chromatography(gradient from 0-40% ethyl acetate+methanol (4/1) in iso-hexanes) toafford 70c (149 mg, 73%) as a white solid after evaporation. ¹H NMR (400MHz, CD₃OD): δ 8.35 (d, J=8.6 Hz, 1H), 8.07-8.01 (m, 1H), 7.93 (d, J=8.5Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 6.98-6.77 (m,2H), 5.48 (q, J=6.5, 6.1 Hz, 1H), 5.20 (d, J=4.0 Hz, 1H), 5.06 (d,J=12.1 Hz, 1H), 4.88 (d, J=12.1 Hz, 1H), 4.25-4.15 (m, 1H), 3.99-3.85(m, 2H), 3.80-3.55 (m, 2H), 3.46 (s, 3H), 2.20-2.09 (m, 1H), 2.07-1.94(m, 1H), 1.92-1.80 (m, 1H), 1.80-1.71 (m, 1H), 1.68-1.64 (m, 6H), 1.59(d, J=7.1 Hz, 3H), 1.54 (s, 9H), 1.37 (d, J=7.0 Hz, 3H), 1.29 (d, J=6.4Hz, 3H). LCMS (m/z) 812.3 [M+H]′ Tr=2.64 min (Gemini 5 u C18 110 Å,50×4.60 mm 5 micron column, 3.5 min, 2 mL/min, 5-100%acetonitrile/water, 0.1% acetic acid modifier gradient).

Compound 70

To 70c (139 mg, 0.17 mmol) in tetrahydrofuran (10 mL) was added asolution of lithium hydroxide (4.5 mg, 0.19 mmol) in water (5 mL). Afterstirring at room temperature for 2 h, 1M hydrochloric acid was added(0.20 mL of 1M solution in water, 0.20 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (5 mL, 20 mmol) at room temperature under argon for 4 hours.Reaction mixture was concentrated under reduced pressure. This residuewas dissolved in N,N-dimethylformamide (5 mL) and the obtained solutionwas added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (98 mg, 0.26 mmol),N,N-diisopropylethylamine (110 mg, 0.86 mmol) and dichloromethane (100mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (50 ml) and brine (50 mL), dried over magnesium sulfate, filteredand concentrated under reduced pressure. The residue was purified byHPLC to afford compound 70 (20 mg, 21%) as a white solid afterlyophilization. ¹H NMR (400 MHz, CDCl₃): δ 9.05 (s, 1H), 8.05 (bs, 1H),7.69 (s, 1H), 7.24 (s, 1H), 7.61 (dd, J=8.4, 1.6 Hz, 1H), 7.43 (d, J=8.5Hz, 1H), 6.62 (d, J=12.3 Hz, 1H), 6.33 (d, J=12.3 Hz, 1H), 6.14 (m, 1H),5.86 (bs, 1H), 5.08 (m, 1H), 4.53 (s, 1H), 4.12 (d, J=6.1 Hz, 1H),3.57-3.49 (m, 2H), 2.02-1.92 (bm, 2H), 1.68 (m, 1H), 1.72-1.45 (cm, 8H),1.07 (s, 9H), 0.91 (m, 3H). LCMS (m/z) 564.3 [M+H]′ Tr=2.42 min.

Example 71 Compound 71

Following the synthetic sequence described in Example 70, the indicatedN-Boc-protected quinolnyl seco-acid was prepared in 5% yield beginningfrom 1 gram commercially available 4,4,4-trifluoro-DL-valine obtainedfrom Apollo Scientific, Inc.

To this compound (139 mg, 0.17 mmol) in tetrahydrofuran (10 mL) wasadded a solution of lithium hydroxide (4.5 mg, 0.19 mmol) in water (5mL). After stirring at room temperature for 2 hour, 1M hydrochloric acidwas added (0.20 mL of 1M solution in water, 0.20 mmol) and the reactionmixture was concentrated under reduced pressure and the residuepartitioned between dichloromethane (50 mL) and water (50 mL). Theorganic layer was collected, concentrated under reduced pressure anddried under high vacuum for one day. The residue was treated with 4Mhydrogen chloride in 1,4-dioxane (5 mL, 20 mmol) at room temperatureunder argon for 4 hours. Reaction mixture was concentrated under reducedpressure. This residue was dissolved in N,N-dimethylformamide (5 mL) andthe obtained solution was added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (98 mg, 0.26 mmol),N,N-diisopropylethylamine (110 mg, 0.86 mmol) and dichloromethane (100mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (50 ml) and brine (50 mL), dried over magnesium sulfate, filteredand concentrated under reduced pressure. The residue was purified byHPLC on C18 column with acetonitrile/water eluent to afford 71 as thelatest eluting of three isomers (7 mg, 8%) as a white solid afterevaporation. ¹H NMR (400 MHz, CD₃CN): δ 7.86 (s, 1H), 7.72 (s, 1H), 7.61(d, J=5.8 Hz, 1H), 7.43 (d, J=5.8 Hz, 1H), 7.31 (m, 1H), 6.50 (d, J=12.2Hz, 1H), 6.34 (d, J=12.2 Hz, 1H), 5.60 (m, 2H), 5.08 (m, 1H), 4.38 (m,1H), 4.18 (d, J=9.1 Hz, 1H), 3.52 (m, 1H), 2.91 (m, 1H), 2.59 (m, 1H),1.94-1.48 (cm, 10H), 1.42 (s, 3H), 1.38 (cm, 5H). LCMS (m/z) 604.3[M+H]′ Tr=2.42 min.

Example 72 Compound 72

To a solution of 2-amino-3-fluoro-3-methyl butanoic acid obtained fromSigma-Aldrich Inc. (1 g, 7.5 mmol) in 1M sulfuric acid (15 ml, 1Maqueous solution), cooled to 0° C., was added a solution of sodiumnitrite (1.0 g, 15 mmol) in water (8 ml). The temperature was maintainedbelow 50° C. during the addition, and the mixture was stirred at suchovernight. The solution was then saturated with ammonium sulfate,extracted with diethyl ether (5×25 ml), dried over sodium sulfate andevaporated under reduced pressure giving the title compound (0.4 g, 37%)as a colorless oil that crystallized on standing. ¹H NMR (400 MHz,CDCl₃): δ 4.34 (d, 1H), 1.35 (d, 6H). LCMS (m/z) 137.4 [M+H], Tr=0.79min.

Compound 72b

A solution of 1d (517 mg, 1.19 mmol) in dichloromethane (20 mL) wascooled in an ice water bath. Trimethylsilyl trifluoromethanesulfonate(398 mg, 1.79 mmol) was added dropwise at 0° C. under argon, and theresulting solution was stirred at room temperature for 30 minutes. Thereaction mixture was evaporated to dryness and the resulting cruderesidue (LCMS (m/z) 332.2/334.3 [M+H]′ Tr=2.06 min) was dissolved inanhydrous dichloromethane (20 mL) and cooled to 0° C. To this was added72a (192 mg, 1.43 mmol), N,N-diisopropylethylamine (0.51 mL, 2.98 mmol),and (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate (633 mg, 1.43 mmol). The reaction was allowed towarm to room temperature and stirred for 8 hours. The mixture wasevaporated to dryness and the residue was dissolved in ethyl acetate(100 mL) and washed with 10% citric acid (100 mL), saturated NaHCO₃ (100mL), and brine (100 mL). The organic layer was dried over MgSO₄, onevolume equivalent of hexane was added and this solution was filteredthrough a 5 cm layer of silica gel, silica gel layer was washed with 50mL of ethyl acetate/hexane mixture (1/1). The desired product was washedout with ethyl acetate (100 mL), concentrated under reduced pressure andco-distilled with dichloromethane. 72b (400 mg, 80%) was isolated afterdrying under high vacuum for one day. ¹H NMR (400 MHz, CD₃OD): δ 5.53(q, J=6.9 Hz, 1H), 5.10 (d, J=12.1 Hz, 1H), 4.92 (d, J=12.2 Hz, 1H),3.96 (d, J=2.6 Hz, 1H), 3.94 (dd, J=7.1, 4.7 Hz, 1H), 3.86 (qd, J=6.4,2.5 Hz, 1H), 3.81-3.60 (m, 2H), 3.42 (s, 3H), 2.34-2.14 (m, 1H),2.09-1.90 (m, 2H), 1.87-1.76 (m, 1H), 1.41 (d, J=6.9 Hz, 3H), 1.32 (d,J=6.4 Hz, 3H). LCMS (m/z) 450.1 [M+H]′ Tr=2.11 min.

Compound 72c

Following the procedure described in Example 70, into an oven-dried,argon purged flask were placed 2-methyl-6-nitrobenzoic anhydride (172mg, 0.5 mmol), 4-dimethylaminopyridine (64 mg, 0.50 mmol), the N-Bocprotected quinoline 1,3-dioxane carboxylic acid 63c (96 mg, 0.25 mmol),and anhydrous dichloromethane (10 mL). Into the resulting solution wasadded N,N-diisopropylethylamine (0.13 mL, 0.75 mmol) and this reactionmixture was stirred at room temperature for 10 minutes. Compound 72b(168 mg, 0.38 mmol) was added via syringe as a solution in anhydrousdichloromethane (10 mL). After stirring for 2 hours at room temperature,the reaction mixture was transferred to a separatory funnel and washedwith water (20 mL, 10 mL of brine was added to support the separation).The aqueous phase was extracted with dichloromethane (20 mL). Combinedorganic extracts were washed with brine (20 mL) and dried over magnesiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by silica gel chromatography (gradient from 0-40% ethylacetate+methanol (4/1) in iso-hexanes) to afford the 72c (149 mg, 73%)as a white solid after evaporation. ¹H NMR (400 MHz, CD₃OD): δ 8.35 (d,J=8.6 Hz, 1H), 8.07-8.01 (m, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.88 (d, J=8.4Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 6.98-6.77 (m, 2H), 5.48 (q, J=6.5, 6.1Hz, 1H), 5.20 (d, J=4.0 Hz, 1H), 5.06 (d, J=12.1 Hz, 1H), 4.88 (d,J=12.1 Hz, 1H), 4.25-4.15 (m, 1H), 3.99-3.85 (m, 2H), 3.80-3.55 (m, 2H),3.46 (s, 3H), 2.20-2.09 (m, 1H), 2.07-1.94 (m, 1H), 1.92-1.80 (m, 1H),1.80-1.71 (m, 1H), 1.68-1.64 (m, 6H), 1.59 (d, J=7.1 Hz, 3H), 1.54 (s,9H), 1.37 (d, J=7.0 Hz, 3H), 1.29 (d, J=6.4 Hz, 3H). LCMS (m/z)860.1/862.5 [M+H].′ Tr=2.64 min.

Compound 72

To 72c (139 mg, 0.17 mmol) in tetrahydrofuran (10 mL) was added asolution of lithium hydroxide (4.5 mg, 0.19 mmol) in water (5 mL). Afterstirring at room temperature for 2 hour, 1M hydrochloric acid was added(0.20 mL of 1M solution in water, 0.20 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (5 mL, 20 mmol) at room temperature under argon for 4 hours.Reaction mixture was concentrated under reduced pressure. This residuewas dissolved in N,N-dimethylformamide (5 mL) and the obtained solutionwas added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (98 mg, 0.26 mmol),N,N-diisopropylethylamine (110 mg, 0.86 mmol) and dichloromethane (100mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (50 ml) and brine (50 mL), dried over magnesium sulfate, filteredand concentrated under reduced pressure. The residue was purified bysilica gel chromatography (gradient from 0-60% ethyl acetate+methanol(4/1) in iso-hexanes) to afford the title compound (32 mg, 33%) as awhite solid after evaporation. ¹H NMR (400 MHz, CD₃OD): δ 8.26 (s, 1H),7.83 (d, J=8.2 Hz, 1H), 7.83 (d, J=8.2 Hz, 1H), 7.51 (m, 2H), 6.71 (d,J=12.3 Hz, 1H), 6.19 (d, J=12.3 Hz, 1H), 5.86 (q, J=7.2 Hz, 1H), 5.48(d, J=8.2 Hz, 1H), 5.08 (m, 2H), 4.41 (m, 1H), 3.77-3.62 (m, 1H), 3.59(m, 1H), 2.68 (m, 1H), 2.31-2.24 (m, 1H), 1.99-1.91 (m, 1H), 1.72-1.54(cm, 8H), 1.42 (s, 3H), LCMS (m/z) 612.1 [M+H]′ Tr=2.22 min.

Example 73 Compound 73

Following the synthetic sequence described in Example 43, instead using1.5 g methyl-oxolane-3-carboxylate (C.A.S. #53662-85-4) supplied byEnamine, Ltd., the indicated bis-protected seco-acid above was preparedin 110 mg quantity (1.2% yield). This intermediate was then carriedthrough the standard multistep synthetic sequence reported in Example 70to afford Compound 73, where 4 mg was obtained after final HPLCpurification as the a diasteromer mixture. ¹H NMR (400 MHz, CD₃CN): δ8.81 (m, 1H), 8.15 (d, J=6.1 Hz, 1H), 7.73 (d, J=6.2 Hz, 1H), 7.60 (m,1H), 7.35 (m, 1H), 7.06 (m, 1H), 6.48-6.33 (m, 2H), 5.62 (m, 1H), 5.08(m, 1H), 4.95 (m, 1H), 4.30 (m, 2H), 4.08 (m, 2H), 3.90-3.69 (cm, 4H),3.56 (m, 2H), 3.39 (m, 2H), 2.68 (cm, 3H), 2.31-2.24 (m, 2H), 1.55-1.35(cm, 7H), 0.92-0.82 (cm, 7H). LCMS (m/z) 578.2 [M+H], Tr=2.11 min.

Examples 74, 75, 76, and 77 Compounds 74, 75, 76, and 77

Intermediate 74a

To a mixture of the bromide 39a (556 mg, 1.583 mmol),ethyl-4-oxo-1-vinylcyclohexanecarboxylate (obtained from SmallMolecules, Inc.), (559 mg, 2.849 mmol), Pd(OAc)₂ (70 mg, 0.312 mmol),and (o-Tol)₃P (100 mg, 0.329 mmol) in 1,4-dioxane (5 mL) was added NEt₃(0.72 mL, 5.166 mmol) and the resulting mixture was heated at 100° C.for 30 min in a microwave reactor. The mixture was concentrated andtreated with water and ethyl acetate (˜50 mL each). After the mixturewas filtered through celite pad, the two fractions in the filtrate wereseparated. After the aqueous fraction was extracted with ethyl acetate(×1), two organic fractions were washed with water (×1), combined, dried(MgSO₄), and concentrated. The residue was purified by CombiFlash usinghexanes-ethyl acetate as eluents to obtain 479 mg (65%) of the product74a. ¹H NMR (400 MHz, Chloroform-d): δ8.07 (d, J=8.3 Hz, 1H), 8.01 (s,1H), 7.75 (d, J=8.5 Hz, 1H), 7.58 (d, J=8.3 Hz, 1H), 7.31 (d, J=8.5 Hz,1H), 6.74 (d, J=16.1 Hz, 1H), 6.47 (d, J=16.3 Hz, 1H), 6.16 (s, 1H),4.27 (q, J=7.1 Hz, 2H), 2.76-2.56 (m, 2H), 2.57-2.38 (m, 4H), 2.18-2.00(m, 2H), 1.47 (s, 9H), 1.32 (t, J=7.1 Hz, 3H). LCMS (m/z) 467.0 [M+H],Tr=2.33 min.

Compound 74b

A solution of 74a (237 mg, 0.508 mmol) in THF (5 mL) was stirred at −60°C. bath as 1.0 M K-selectride in THF (0.61 mL) was added. After thereaction mixture was slowly warmed to 0° C. over 30 min, the mixture wasdiluted with ethyl acetate and washed with water (×2). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 192 mg(81%) 74b as a ˜4:1 mixture. LCMS (m/z) 469.0 [M+H], Tr=2.20 min.

Compound 74c

A mixture of 74b (192 mg, 0.410 mmol) and LiOH (87.7 mg, 2.090 mmol) inTHF (2 mL), MeOH (2 mL), and water (2 mL) was stirred at 50° C. bath for3 h. The mixture was concentrated to ˜1/3 volume, diluted with water,neutralized with 1 N HCl (2.2 mL), and extracted with ethyl acetate(×2). The extracts were washed with water (×1), combined, dried (MgSO₄),and concentrated. A solution of the residue, imidazole (115 mg, 1.689mmol), and TBSCI (79 mg, 0.524 mmol) in DMF was stirred at 35° C. bathfor 1.5 h before addition of additional imidazole (102 mg, 1.498 mmol),and TBSCI (77 mg, 0.511 mmol) were added. The resulting mixture wasstirred at 35° C. for 17 h, concentrated, and diluted with ethyl acetatebefore washing with 5% LiCl solution (×2). After the aqueous fractionswere extracted with ethyl acetate (×1), two organic fractions werecombined, dried (MgSO₄), and concentrated. The residue was dissolved inTHF (2 mL), MeOH (2 mL), and water (2 mL), and stirred with K₂CO₃ (226mg, 1.635 mmol) at rt for 30 min. The solution was concentrated to ˜1/2volume, diluted with water, and acidified with 1 N HCl (˜3.5 mL) beforeextraction with ethyl acetate (×2). The extracts were washed with water(×1), combined, dried (MgSO₄), and concentrated. The residue waspurified by CombiFlash using hexanes-ethyl acetate as eluents to obtain193 mg (85%) of the product 74c as an apparent ˜4:1 isomer mixture. LCMS(m/z) 555.1 [M+H], Tr=2.60 min.

Compound 74d

A mixture of the acid 74c (131 mg, 0.236 mmol), the compound 1e (125 mg,0.289 mmol), Shiina reagent (98 mg, 0.285 mmol), and DMAP (72 mg, 0.589mmol) was dissolved in CH₂Cl₂ (3 mL) at rt. After 3 min, DIPEA (0.1 mL,0.574 mmol) was added and the resulting mixture was stirred at rt for16.5 h. The mixture was diluted with ethyl acetate, and washed withwater (×2). After the aqueous fractions were extracted with ethylacetate (×1), two organic fractions were combined, dried (MgSO₄), andconcentrated. The residue was purified by CombiFlash using hexanes-ethylacetate as eluents to obtain 90 mg (39%) of 74d as a mixture. LCMS (m/z)968.2 [M+H], Tr=3.04 min.

Compounds 74, 75, 76, and 77

A mixture of 74d (90 mg, 0.093 mmol) in THF (1 mL), MeOH (1 mL), andwater (1 mL) was added K₂CO₃ (89 mg, 0.644 mmol) at 0° C. The mixturewas stirred at 0° C. for 2.5 h, 64 h at freezer, at rt for 3 h, and at30° C. for 1.5 h. The mixture was diluted with saturated NaCl solutionand extracted with ethyl acetate (×2). After the aqueous fractions wereextracted with ethyl acetate (×1), two organic fractions were combined,dried (MgSO₄), concentrated, and dried in vacuum. The residue wasdissolved in 4 N HCl in dioxane (3 mL) and stirred at rt for 45 minbefore concentration and drying in vacuum for 2 h to obtain the crudehydroxy amino acid. A suspension of HATU (177 mg, 0.466 mmol) in CH₂Cl₂(100 mL) was stirred at rt as a solution of the crude hydroxylamino acidand DIPEA (0.13 mL, 0.746 mmol) in DMF (7.5 mL) was added over 5 min.After 1.5 h at rt, the mixture was concentrated and the residue wasdiluted with ethyl acetate before washing with 5% LiCl solution (×2),0.1 N HCl (×1), and water (×1). After the aqueous fractions wereextracted with ethyl acetate (×1), two organic fractions were combined,dried (MgSO₄), and concentrated. The residue was separated bypreparative HPLC using a 21.2×250 mm 10 micron C18 Phenomenex Geminisemi-preparative column and acetonitrile/water (containing 0.1% TFAmodifier) as mobile phase at a flow rate of 20 mL/min with a 50 minutegradient consisting of 0 min-5 min: 20% acetonitrile; 5 min-48 min: 20%acetonitrile to 80% acetonitrile; 48 min-50 min: 80% acetonitrile toobtain four separate product fractions. Each fraction was thenseparately concentrated to remove MeCN, neutralized with NaHCO₃solution, diluted with water, and extracted with ethyl acetate (×2). Theseparated extracts were concentrated to obtain 4.6 mg (8%) of 74, 1.1 mg(2%) of 75, 10.5 mg (19%) of 76, and 2.3 mg (4%) of 77.

Compounds 74-77 represent stereoisomers of the compound:

Contemplated stereoisomers include:

Compound 74: First eluting isomer. ¹H NMR (400 MHz, Methanol-d₄): δ8.23(d, J=8.5 Hz, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.77 (d, J=1.7 Hz, 1H), 7.62(dd, J=8.4, 1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.61 (d, J=16.4 Hz,1H), 6.29 (d, J=16.4 Hz, 1H), 5.79 (q, J=7.2 Hz, 1H), 5.23 (d, J=8.6 Hz,1H), 5.06 (q, J=6.6 Hz, 1H), 4.41 (dd, J=13.2, 4.3 Hz, 1H), 3.88 (s,1H), 3.57 (dd, J=11.7, 2.8 Hz, 1H), 2.68 (td, J=13.0, 3.3 Hz, 1H), 2.25(d, J=11.2 Hz, 1H), 2.21-2.06 (m, 3H), 2.00-1.88 (m, 3H), 1.84-1.63 (m,5H), 1.60 (d, J=7.3 Hz, 3H), 1.57 (d, J=6.8 Hz, 3H), 1.61-1.47 (m, 1H),1.28 (s, 1H), 1.05 (d, J=6.8 Hz, 3H), 1.00 (d, J=6.7 Hz, 3H). LCMS (m/z)606.3 [M+H], Tr=2.03 min.

Compound 75: Second eluting isomer. ¹H NMR (400 MHz, Methanol-d₄): δ8.23(d, J=8.5 Hz, 1H), 7.81 (d, J=8.5 Hz, 1H), 7.76 (d, J=1.7 Hz, 1H), 7.60(dd, J=8.4, 1.6 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H), 6.53 (d, J=16.3 Hz,1H), 6.25 (d, J=16.3 Hz, 1H), 5.80 (q, J=7.2 Hz, 1H), 5.28 (d, J=8.6 Hz,1H), 5.06 (q, J=6.7 Hz, 1H), 4.41 (d, J=13.8 Hz, 1H), 3.60 (m, 2H),2.76-2.62 (m, 1H), 2.57 (d, J=14.1 Hz, 1H), 2.33 (dd, J=13.0, 3.9 Hz,1H), 2.29-2.10 (m, 2H), 1.94 (m, 3H), 1.78-1.38 (m, 3H), 1.61 (d, J=7.3Hz, 3H), 1.57 (d, J=6.7 Hz, 3H), 1.28 (s, 4H), 1.07 (d, J=6.8 Hz, 3H),1.04 d, J=6.8 Hz, 3H). LCMS (m/z) 606.3 [M+H], Tr=2.02 min.

Compound 76: Third eluting isomer. ¹H NMR (400 MHz, Methanol-d₄): δ 9.43(d, J=6.2 Hz, 1H), 8.24 (d, J=8.5 Hz, 1H), 8.02-7.94 (m, 1H), 7.84 (d,J=8.4 Hz, 1H), 7.57 (dd, J=8.4, 1.6 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H),6.86 (d, J=16.3 Hz, 1H), 6.33 (d, J=16.3 Hz, 1H), 5.85 (q, J=7.2 Hz,1H), 5.19 (d, J=9.0 Hz, 1H), 5.15 (m, 1H), 4.49-4.38 (m, 1H), 3.83 (s,1H), 3.41 (dd, J=11.8, 2.8 Hz, 1H), 2.79 (td, J=12.8, 3.2 Hz, 1H), 2.39(dd, J=13.2, 3.4 Hz, 1H), 2.21 (m, 1H), 2.18-2.06 (m, 3H), 2.06-1.91 (m,2H), 1.76 (m, 2H), 1.68 (m, 2H), 1.44-1.61 (m, 1H), 1.56 (d, J=6.7 Hz,3H), 1.28 (s, 1H), 1.22 (d, J=7.2 Hz, 3H), 1.04 (d, J=6.9 Hz, 3H), 1.02(d, J=6.7 Hz, 3H). LCMS (m/z) 606.3 [M+H], Tr=2.10 min.

Compound 77: Fourth eluting isomer. ¹H NMR (400 MHz, Methanol-d₄): δ9.45 (d, J=6.3 Hz, 1H), 8.25 (d, J=8.4 Hz, 1H), 8.00 (d, J=1.6 Hz, 1H),7.84 (d, J=8.4 Hz, 1H), 7.55 (dd, J=8.4, 1.6 Hz, 1H), 7.44 (d, J=8.5 Hz,1H), 6.78 (d, J=16.3 Hz, 1H), 6.36 (d, J=16.3 Hz, 1H), 5.88 (q, J=7.2Hz, 1H), 5.45 (d, J=12.0 Hz, 1H), 5.24 (d, J=8.9 Hz, 1H), 5.17 (q, J=6.6Hz, 1H), 4.51-4.39 (m, 1H), 3.71-3.54 (m, 1H), 3.51-3.38 (m, 1H),2.85-2.72 (m, 1H), 2.63 (d, J=13.5 Hz, 1H), 2.43 (m, 2H), 2.24 (m, 1H),1.98 (m, 3H), 1.77-1.59 (m, 2H), 1.57 (d, J=6.7 Hz, 3H), 1.54 (d, J=3.6Hz, 1H), 1.41 (q, J=13.0 Hz, 1H), 1.35-1.26 (m, 2H), 1.26 (d, J=7.2 Hz,3H), 1.07 (d, J=6.8 Hz, 3H), 1.06 (d, J=6.8 Hz, 3H). LCMS (m/z) 606.3[M+H], Tr=2.11 min.

Examples 78 and Example 79 Compounds 78 and 79

Compound 78a

A solution of 50% ethyl glyoxalate in toluene (5.15 mL, 26 mmol) in THF(50 mL) was stirred at −20-−30° C. bath as 0.5 M cyclopropylmagnesiumbromide in THF (50 mL) was added over 10 min. After 2 h at −30° C. bath,the reaction mixture was quenched with water (250 mL) and extracted withethyl acetate (×3). The extracts were washed with water (×1), combined,dried (MgSO₄), and concentrated. The residue was purified by CombiFlashusing hexanes-ethyl acetate as eluents to obtain 882 mg (17%) of 78awith 72% purity. ¹H NMR (400 MHz, Chloroform-d): δ 4.27 (q, J=7.2 Hz,2H), 3.77 (d, J=6.6 Hz, 1H), 1.56 (s, 1H), 1.32 (t, J=7.1 Hz, 3H),1.15-1.03 (m, 1H), 0.59-0.46 (m, 3H), 0.46-0.38 (m, 1H).

Compound 78b

A solution of the indicated acid (100 mg, 0.260 mmol), the alcohol (75mg, 0.375 mmol, 72% purity), and DMAP (79 mg, 0.647 mmol) in CH₂Cl₂ (5mL) were added Shiina reagent (180 mg, 0.523 mmol) and DIEA (0.24 mL,1.378 mmol) at rt. After 16.5 h at rt, the mixture was concentrated andthe residue was dissolved in ethyl acetate before washing with dilutecitric acid solution (×1) and NaHCO₃ solution (×1). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 76 mg(57%) of the product 78b as a diasteromer mixture. ¹H NMR (400 MHz,Chloroform-d): δ8.05 (d, J=8.4 Hz, 1H), 7.99 (s, 1H), 7.72 (d, J=8.4 Hz,1H), 7.61 (dd, J=8.5, 1.7 Hz, 1H), 7.28 (d, J=8.6 Hz, 1H), 6.69 (d,J=16.3 Hz, 1H), 6.63 (d, J=16.2 Hz, 1H), 6.22 (s, 1H), 5.09-4.87 (m,1H), 4.49 (d, J=7.9 Hz, 1H), 4.21 (p, J=7.0 Hz, 2H), 1.53 (d, J=7.3 Hz,3H), 1.51 (s, 3H), 1.49 (s, 3H), 1.47 (s, 9H), 1.30 (m, 1H), 1.26 (t,J=7.1 Hz, 3H), 0.72-0.52 (m, 3H), 0.51-0.41 (m, 1H). LCMS (m/z) 511.0[M+H], Tr=2.49 min.

Compound 78c

To a solution of the ethyl ester reactant 78b (76 mg, 0.149 mmol) in THF(2 mL), MeOH (2 mL), and water (2 mL) was added K₂CO₃ (104 mg, 0.752mmol) at 0° C. After stirring at 0° C. for 1.5 h and at rt for 10.5 h,the mixture was acidified using 1 N HCl (1.6 mL), diluted with water,and extracted with ethyl acetate (×2). After the aqueous fractions wereextracted with ethyl acetate (×1), two organic fractions were combined,dried (MgSO₄), and concentrated. The residue was purified by CombiFlashusing hexanes-ethyl acetate as eluents to obtain 49 mg (68%) of theintermediate product with ˜15% of impurity. LCMS (m/z) 483.0 [M+H]. Asolution of the dipeptide (48 mg, 0.111 mmol) in 4 N HCl in dioxane (2mL) was stirred at rt for 1 h. After the mixture was concentrated andthe residue was co-evaporated with toluene, A solution of the residue,the previously obtained acid (49 mg), and HATU (80 mg, 0.210 mmol) inCH₂Cl₂ (3 mL) and DMF (1 mL) was stirred at rt as DIEA (0.1 mL, 0.574mmol) was added. After 30 min at rt, the mixture was diluted with EA andwashed with 5% LiCl solution (×2), citric acid solution (×1), saturatedNaHCO₃, (×1), and saturated NaCl solution (×1). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 64 mg(79%) of the product 78c as a diasteromer mix. ¹H NMR (400 MHz,Chloroform-d): δ8.05 (d, J=8.4 Hz, 1H), 8.00 (s, 1H), 7.71 (d, J=8.3 Hz,1H), 7.67-7.60 (m, 1H), 7.28 (d, J=8.5 Hz, 1H), 7.03 (dd, J=16.5, 7.6Hz, 1H), 6.71 (dd, J=16.2, 3.8 Hz, 1H), 6.64 (d, J=16.2 Hz, 1H), 6.21(s, 1H), 5.37-5.20 (m, 1H), 4.97 (s, 1H), 4.92 (d, J=11.9 Hz, 1H), 4.77(dd, J=7.8, 3.2 Hz, 1H), 4.69 (dd, J=11.9, 3.7 Hz, 1H), 4.36-4.18 (m,1H), 3.81 (t, J=8.9 Hz, 1H), 3.65 (m, 1H), 2.84 (br, 1H), 2.14 (m, 1H),1.88 (m, 1H), 1.77-1.59 (m, 2H), 1.55 (s, 3H), 1.53 (s, 3H), 1.51 (d,J=8.2 Hz, 3H), 1.46 (s, 9H), 1.24 (m, 4H), 0.65 (m, 1H), 0.50 (m, 3H).LCMS (m/z) 796.0 [M+H], Tr=2.50 min.

Compound 78 and Compound 79

A mixture of the reactant (64 mg, 0.080 mmol) in THF (1 mL) and water (1mL) was stirred at 0° C. as 1N KOH (0.09 mL) was added. After stirringat 0° C. for 1.25 h, the mixture was acidified using 1 N HCl (0.1 mL),diluted with water, and extracted with ethyl acetate (×2). After theaqueous fractions were extracted with ethyl acetate (×1), two organicfractions were combined, dried (MgSO₄), and concentrated. The residuewas dissolved in 4 N HCl in dioxane (2 mL) and stirred at rt for 1 h.After the mixture was concentrated and the residue was co-evaporatedwith toluene, a solution of the residue and HATU (155 mg, 0.408 mmol) inDMF (2.5 mL) and CH₂Cl₂ (25 mL) was stirred at 0° C. as DIEA (0.12 mL,0.689 mmol) was added. After 30 min, the solution was concentrated,diluted with ethyl acetate before washing with 5% LiCl solution (×2),citric acid solution (×1), and saturated NaHCO₃ solution (×1). After theaqueous fractions were extracted with ethyl acetate (×1), two organicfractions were combined, dried (MgSO₄), and concentrated. The residuewas separated by preparative HPLC to obtain 2 fractions. Each fractionwas separately concentrated to remove MeCN, neutralized with NaHCO₃solution, diluted with water, and extracted with ethyl acetate (×2). Theextracts were combined, dried (MgSO₄), and concentrated to obtain 11.9mg (23%) of 78 and 11.0 mg (22%) of 79.

Compound 78: ¹H NMR (400 MHz, Methanol-d₄): δ8.63 (d, J=7.2 Hz, 1H),8.23 (d, J=8.5 Hz, 1H), 7.85 (s, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.57 (dd,J=8.5, 1.7 Hz, 1H), 7.42 (d, J=8.5 Hz, 1H), 6.56 (d, J=16.2 Hz, 1H),6.35 (d, J=16.2 Hz, 1H), 5.87-5.72 (m, 1H), 5.14-5.01 (m, 1H), 4.89 (d,J=8.9 Hz, 1H), 4.43 (dd, J=13.8, 4.4 Hz, 1H), 3.57 (dd, J=11.8, 2.9 Hz,1H), 2.69 (td, J=13.0, 3.3 Hz, 1H), 2.34-2.21 (m, 1H), 1.94 (d, J=13.4Hz, 1H), 1.76-1.61 (m, 1H), 1.59 (d, J=7.2 Hz, 3H), 1.56 (d, J=6.7 Hz,3H), 1.51 (s, 3H), 1.40 (s, 3H), 1.30 (m, 2H), 0.69-0.57 (m, 3H),0.56-0.42 (m, 1H). LCMS (m/z) 548.3 [M+H], Tr=2.25 min.

Compound 79: ¹H NMR (400 MHz, Methanol-d₄): δ9.27 (d, J=6.2 Hz, 1H),8.24 (s, 1H), 8.21 (d, J=8.5 Hz, 1H), 8.02 (d, J=6.7 Hz, 1H), 7.80 (d,J=8.3 Hz, 1H), 7.46 (dd, J=8.3, 1.7 Hz, 1H), 7.41 (d, J=8.4 Hz, 1H),6.72 (d, J=16.0 Hz, 1H), 6.57 (d, J=16.1 Hz, 1H), 5.91-5.76 (m, 1H),5.13 (m, 1H), 4.73 (d, J=8.9 Hz, 1H), 4.43 (dd, J=8.8, 4.0 Hz, 1H), 3.55(dd, J=11.9, 2.9 Hz, 1H), 2.69 (td, J=13.1, 3.3 Hz, 1H), 2.32 (dd,J=13.4, 3.4 Hz, 1H), 1.93 (d, J=13.2 Hz, 1H), 1.68 (qt, J=13.4, 4.4 Hz,1H), 1.57 (d, J=6.9, Hz, 3H), 1.56 (d, J=6.9, Hz, 3H), 1.54 (s, 3H),1.53-1.45 (m, 1H), 1.42 (s, 3H), 1.28 (s, 1H), 0.78-0.59 (m, 3H), 0.50(m, 1H). LCMS (m/z) 548.3 [M+H], Tr=2.33 min.

Example 80 Compound 80

Compound 80

To a solution of 74 (7.13 mg, 0.012 mmol) in pyridine (0.15 mL) wasadded isobutyryl chloride (0.05 mL) at rt. After 1 h at rt, the mixturewas diluted with ethyl acetate, washed with citric acid solution (×1)and saturated NaHCO₃ solution (×1). After the aqueous fractions wereextracted with EA (×1), two organic fractions were combined, dried(MgSO₄), and concentrated. The residue was purified by CombiFlash usinghexanes-ethyl acetate as eluents to obtain 6.4 mg (80%) of the product80. ¹H NMR (400 MHz, Methanol-d₄): δ8.83 (d, J=7.5 Hz, 1H), 8.23 (d,J=8.5 Hz, 1H), 7.83 (d, J=8.5 Hz, 1H), 7.79 (s, 1H), 7.64 (dd, J=8.4,1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.61 (d, J=16.4 Hz, 1H), 6.30 (d,J=16.4 Hz, 1H), 5.80 (q, J=7.1 Hz, 1H), 5.26 (d, J=8.5 Hz, 1H), 5.07 (q,J=6.7 Hz, 1H), 4.96 (s, 1H), 4.41 (dd, J=13.5, 4.1 Hz, 1H), 3.62-3.54(m, 1H), 2.68 (td, J=13.2, 3.4 Hz, 1H), 2.60 (p, J=7.0 Hz, 1H), 2.33 (d,J=13.4 Hz, 1H), 2.25 (d, J=12.0 Hz, 1H), 2.21-2.09 (m, 2H), 2.05-1.76(m, 7H), 1.76-1.63 (m, 1H), 1.60 (d, J=7.2 Hz, 3H), 1.57 (d, J=6.7 Hz,3H), 1.56-1.46 (m, 1H), 1.20 (d, J=7.0 Hz, 6H), 1.06 (d, J=6.8 Hz, 3H),1.02 (d, J=6.7 Hz, 3H). LCMS (m/z) 676.3 [M+H], Tr=1.35 min.

Example 81 Compound 81

Compound 81

To a solution of the 75 (6.6 mg, 0.011 mmol) in pyridine (0.15 mL) wasadded isobutyryl chloride (0.05 mL) at rt. After 1 h at rt, the mixturewas diluted with ethyl acetate, washed with citric acid solution (×1)and saturated NaHCO₃ solution (×1). After the aqueous fractions wereextracted with ethyl acetate (×1), two organic fractions were combined,dried (MgSO₄), and concentrated. The residue was purified by CombiFlashusing hexanes-ethyl acetate as eluents to obtain 5.1 mg (69%) of theproduct. ¹H NMR (400 MHz, Methanol-d₄): δ8.27 (d, J=8.5 Hz, 1H), 7.83(d, J=8.2 Hz, 1H), 7.79 (s, 1H), 7.64 (d, J=8.3 Hz, 1H), 7.46 (d, J=8.5Hz, 1H), 6.58 (d, J=16.3 Hz, 1H), 6.29 (d, J=16.4 Hz, 1H), 5.81 (p,J=7.4 Hz, 1H), 5.28 (d, J=8.4 Hz, 1H), 5.08 (q, J=6.8 Hz, 1H), 4.77 (m,1H), 4.42 (d, J=15.0 Hz, 1H), 3.58 (d, J=11.6, 3.2 Hz, 1H), 2.68 (td,J=12.8, 3.2 Hz, 1H), 2.56 (m, 1H), 2.52 (hept, J=7.0 Hz, 1H), 2.35 (d,J=13.1 Hz, 1H), 2.30-2.11 (m, 2H), 2.10-1.88 (m, 3H), 1.75 (td, J=13.6,2.8 Hz, 1H), 1.71-1.40 (m, 5H), 1.61 (d, J=7.5 Hz, 3H), 1.59 (d, J=6.9Hz, 3H), 1.14 (d, J=7.0, Hz, 6H), 1.08 (d, J=6.8 Hz, 3H), 1.03 (d, J=6.7Hz, 3H). LCMS (m/z) 676.5 [M+H], Tr=1.20 min.

Example 82 Compound 82

Compound 82

To a solution of the 75 (7.0 mg, 0.012 mmol) in pyridine (0.15 mL) wasadded n-butyryl chloride (0.05 mL) at rt. After 1 h at rt, the mixturewas diluted with ethyl acetate, washed with 10% citric acid solution(×1) and saturated NaHCO₃ solution (×1). After the aqueous fractionswere extracted with ethyl acetate (×1), two organic fractions werecombined, dried (MgSO₄), and concentrated. The residue was purified byCombiFlash using hexanes-ethyl acetate as eluents to obtain 5.7 mg (73%)of the product 82. ¹H NMR (400 MHz, Methanol-d₄): δ8.23 (d, J=8.5 Hz,1H), 7.82 (d, J=8.5 Hz, 1H), 7.77 (s, 1H), 7.61 (dd, J=8.4, 1.7 Hz, 1H),7.43 (d, J=8.5 Hz, 1H), 6.56 (d, J=16.4 Hz, 1H), 6.26 (d, J=16.3 Hz,1H), 5.80 (q, J=7.2 Hz, 1H), 5.28 (d, J=8.6 Hz, 1H), 5.06 (q, J=6.6 Hz,1H), 4.77 (m, 1H), 4.47-4.34 (m, 1H), 3.58 (dd, J=12.8, 2.8 Hz, 1H),2.68 (td, J=12.8, 3.2 Hz, 1H), 2.58 (d, J=13.9 Hz, 1H), 2.41-2.31 (m,1H), 2.28 (t, J=7.3 Hz, 2H), 2.24-2.11 (m, 1H), 2.10-1.88 (m, 3H),1.80-1.38 (m, 9H), 1.61 (d, J=7.3 Hz, 3H), 1.57 (d, J=6.8 Hz, 3H), 1.07(d, J=6.8 Hz, 3H), 1.03 (d, J=6.7 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H). LCMS(m/z) 676.4 [M+H], Tr=1.19 min.

Example 83 Compound 83

Compound 83a

To a mixture of the [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester 39a (1160 mg, 3.303 mmol), the vinyl acid (833 mg,4.953 mmol), Pd(OAc)₂ (149 mg, 0.664 mmol), and (o-Tol)₃P (203 mg, 0.667mmol) in dioxane (18 mL) was added NEt₃ (1.65 mL, 11.838 mmol) and theresulting mixture was heated at 100° C. for 30 min in a microwavereactor. The mixture was filtered through celite pad, and the filtratewas diluted with ethyl acetate before washing with water (×2). After theaqueous fractions were extracted with ethyl acetate (×1), two organicfractions were washed with water (×1), combined, dried (MgSO₄), andconcentrated. The residue was purified by CombiFlash using hexanes-ethylacetate as eluents to obtain 1033 mg (71%) of the product 83a. ¹H NMR(400 MHz, Chloroform-d): δ 8.23 (s, 1H), 8.14 (br d, 1H), 7.76 (d, J=8.5Hz, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.39 (br, 1H), 6.87 (d, J=16.3 Hz, 1H),6.60 (d, J=16.2 Hz, 1H), 6.49 (s, 1H), 5.04 (s, 1H), 2.81-2.58 (m, 4H),2.49 (m, 2H), 2.13 (m, 2H), 1.55 (br, 3H), 1.40 (s, 9H). LCMS (m/z)439.0 [M+H], Tr=0.87 min.

Compound 83b

A solution of the 83a (150 mg, 0.342 mmol) in MeOH (2 mL) and water (2mL) was stirred at rt as methoxylamine hydrochloride (70 mg, 0.838 mmol)and 1 N KOH (0.7 mL) were added. After 1 h at rt, the mixture wasdiluted with ethyl acetate and washed with 10% citric acid (×1) andwater (×1). After the aqueous fractions were extracted with ethylacetate (×1), two organic fractions were washed with water (×1),combined, dried (MgSO₄), and concentrated. The residue was purified byCombiFlash using hexanes-EA as eluents to obtain 144 mg (90%) of theproduct 83b. LCMS (m/z) 468.0 [M+H], Tr=0.98 min.

Compound 83c

A mixture of the acid 83b (144 mg, 0.308 mmol), 1e (162 mg, 0.374 mmol),Shiina reagent (276 mg, 0.802 mmol), and DMAP (107 mg, 0.876 mmol) wasdissolved in CH₂Cl₂ (5 mL) at rt. After 3 min, DIEA (0.15 mL, 0.861mmol) was added and the resulting mixture was stirred at rt for 16 h.The mixture was diluted with ethyl acetate, and washed with water (×2).After the aqueous fractions were extracted with ethyl acetate (×1), twoorganic fractions were combined, dried (MgSO₄), and concentrated. Theresidue was purified by CombiFlash using

hexanes-ethyl acetate as eluents to obtain 161 mg (59%) of the product83c as an isomer mixture. LCMS (m/z) 881.3 [M+H], Tr=1.37 min.

Compound 83

A mixture of the 83c (161 mg, 0.182 mmol) in THF (2 mL) and water (2 mL)was stirred at 0° C. as 1 N KOH (0.19 mL) was added. After stirring at0° C. for 1 h, the mixture was acidified using 1 N HCl (0.20 mL),diluted with water, and extracted with ethyl acetate (×2). After theaqueous fractions were extracted with ethyl acetate (×1), two organicfractions were combined, dried (MgSO₄), and concentrated. The residuewas stirred in CH₂Cl₂ (2 mL) and 4 N HCl in dioxane (2 mL) at rt for 1h. After the mixture was concentrated and the residue was co-evaporatedwith toluene (×2), a solution of the residue and HATU (150 mg, 0.394mmol) in DMF (2.5 mL) and CH₂Cl₂ (50 mL) was stirred at 0° C. as DIEA(0.2 mL, 1.148 mmol) was added. After 1.5 h at 0° C., the solution wasconcentrated, diluted with ethyl acetate before washing with 5% LiClsolution (×2). After the aqueous fractions were extracted with ethylacetate (×1), two organic fractions were combined, dried (MgSO₄), andconcentrated. The residue was purified by CombiFlash using hexanes-ethylacetate as eluents. The partially purified product was further purifiedby preparative HPLC to obtain 17.2 mg (23%) of compound 83 as oneisomer. ¹H NMR (400 MHz, Methanol-d₄): δ9.40 (d, J=5.9 Hz, 1H), 8.87 (d,J=7.5 Hz, 1H), 8.22 (d, J=8.5 Hz, 1H), 7.81 (d, J=8.5 Hz, 1H), 7.78 (s,1H), 7.61 (dd, J=8.5, 1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.60 (d,J=16.4 Hz, 1H), 6.29 (d, J=16.4 Hz, 1H), 5.81 (q, J=7.2 Hz, 1H), 5.30(d, J=8.6 Hz, 1H), 5.12-5.01 (m, 1H), 4.40 (dd, J=13.8, 4.2 Hz, 1H),3.79 (s, 3H), 3.56 (dd, J=11.6, 2.9 Hz, 1H), 3.18 (dt, J=15.0, 4.3 Hz,1H), 2.68 (td, J=13.0, 3.2 Hz, 1H), 2.61-2.50 (m, 1H), 2.48-2.11 (m,5H), 2.06-1.86 (m, 2H), 1.83-1.62 (m, 3H), 1.60 (d, J=7.3 Hz, 3H), 1.57(d, J=6.8 Hz, 3H), 1.51 (dd, J=13.3, 9.6 Hz, 1H), 1.06 (d, J=6.8 Hz,3H), 1.01 (d, J=6.6 Hz, 3H). LCMS (m/z) 633.5 [M+H], Tr=1.05 min.

Example 84 Compound 84

Compound 84a

1-tert-butyl 4-ethyl 4-vinylpiperidine-1,4-dicarboxylate 84a wasprepared according to the reported method for enolate arylation reportedby Bercot, E. A. et al. in Organic Letters, 2008, 10, p. 5251-5254 using1-tert-butyl 4-ethyl piperidine-1,4-dicarboxylate in 12% yield.

¹H NMR (400 MHz, Chloroform-d): δ 5.76 (dd, J=17.5, 10.7 Hz, 1H),5.19-5.02 (m, 2H), 4.14 (q, J=7.1 Hz, 2H), 3.80-3.59 (m, 2H), 3.02 (ddd,J=13.8, 10.2, 3.3 Hz, 2H), 2.10 (dtt, J=13.6, 3.2, 1.6 Hz, 2H), 1.56(ddd, J=13.7, 10.1, 4.1 Hz, 2H), 1.41 (s, 9H), 1.22 (t, J=7.1 Hz, 3H).LCMS [M+H]⁺=184.05 (-Boc). Tr=2.43 min

Compound 84

1-tert-butyl 4-ethyl piperidine-1,4-dicarboxylate (282 mg) washydrolyzed to its acid, and this was then followed by the Heck reactionand subsequent Shiina reaction to form 84 (129 mg) following the methodreported in Example 36. ¹H NMR (400 MHz, Methanol-d₄): δ 9.41 (d, J=6.2Hz, 1H), 8.28-8.18 (m, 1H), 8.00 (s, 1H), 7.57 (d, J=8.5 Hz, 1H), 7.43(d, J=8.5 Hz, 1H), 6.84 (d, J=16.3 Hz, 1H), 6.34 (d, J=16.3 Hz, 1H),5.86 (q, J=7.3 Hz, 1H), 5.44 (d, J=12.2 Hz, 1H), 5.23 (d, J=8.9 Hz, 1H),5.15 (t, J=6.4 Hz, 1H), 4.43 (d, J=13.2 Hz, 1H), 3.96-3.80 (m, 2H), 2.79(d, J=12.7 Hz, 1H), 2.47-2.15 (m, 4H), 1.95 (m, 1H), 1.80 (m, 2H), 1.55(d, J=6.7 Hz, 4H), 1.46 (s, 9H), 1.32-1.16 (m, 10H), 1.04 (dd, J=8.3,6.7 Hz, 6H). LCMS [M+H]⁺=691.30; Tr=2.02 min.

Example 85 Compound 85

Compound 85a

Following the procedure reported in Example 84, the N-Boc group of1-tert-butyl 4-ethyl 4-vinylpiperidine-1,4-dicarboxylate 84a (450 mg)was removed via treatment with 4N HCl in dioxane/DCM at RT for 1 h togive ethyl 4-vinylpiperidine-4-carboxylate. Ethyl4-vinylpiperidine-4-carboxylate reacted with MsCl with TEA in DCM at RTfor 18 h to give ethyl1-(methylsulfonyl)-4-vinylpiperidine-4-carboxylate 85a (409 mg, 98%yield). LCMS [M+H]⁺=249.90 (-Boc); Tr=1.96 min.

Compound 85

Compound 85a (409 mg) was hydrolyzed to its acid, and the materialsubmitted Heck coupling, and Shiina-mediated lactonization reaction wasused to form 85 (34 mg) in a manner like that reported in Example 84. ¹HNMR (400 MHz, Methanol-d₄): δ 8.22 (d, J=8.5 Hz, 1H), 7.85-7.74 (m, 2H),7.70-7.58 (m, 1H), 7.42 (d, J=8.5 Hz, 1H), 6.45 (dd, J=132.8, 16.3 Hz,2H), 5.79 (q, J=7.1 Hz, 1H), 5.28 (d, J=8.5 Hz, 1H), 5.05 (q, J=6.5 Hz,1H), 4.39 (d, J=13.2 Hz, 1H), 4.06 (s, 3H), 3.70 (dd, J=18.6, 5.4 Hz,1H), 3.64 (s, 2H), 3.60-3.51 (m, 1H), 3.04-2.84 (m, 2H), 2.73-2.60 (m,1H), 2.53 (d, J=13.7 Hz, 1H), 2.34 (d, J=13.7 Hz, 1H), 2.30-2.09 (m,2H), 2.04-1.82 (m, 3H), 1.64 (dd, J=10.8, 6.5 Hz, 1H), 1.57 (dd, J=10.6,7.0 Hz, 6H), 1.49 (dd, J=9.7, 5.2 Hz, 1H), 1.45-1.18 (m, 2H), 1.02 (dd,J=17.8, 6.7 Hz, 6H). LCMS [M+H]⁺=669.49; Tr=0.95 min.

Example 86 and Example 87 Compounds 86 and 87

Compound 86a

The reactant t-butyl-L-threonine (supplied by Sigma-Aldrich, Inc.) (1 g,5.7 mmol) in the solution of water (7 mL)/acetone (14 mL)/sulfuric acid(7.2 mL) was cooled to 0° C. A solution of sodium nitrite (1.18 g, 17.1mmol) in water (5 mL) was added dropwise. The mixture was stirred andwarmed to rt overnight. After acetone was removed via rotaryevaporation, the residue was extracted with EtOAc twice and washed withbrine. The organic layer was dried through (MgSO₄), and concentrated.The residue purified by CombiFlash on silica gel column using MeOH/DCM.It gave 674 mg (67% yield) of (2S,3R)-3-tert-butoxy-2-hydroxybutanoicacid 86a. ¹H NMR (400 MHz, Chloroform-d): δ 4.18-4.02 (m, 1H), 2.40 (m,1H), 1.32-1.25 (m, 12H).

Compound 86b

(2S,3R)-3-tert-butoxy-2-hydroxybutanoic acid 86a (337 mg, 1.9 mmol) wasreacted with the indicated dipeptide resulting from N-Boc removal from1d as described in the synthesis of 1e (1.2 eq) with DIPEA (2 mL), BOP(924 mg, 2.09 mmol) in DMF at 0° C. for 30 min, and then at RT for 30min. It was quenched with 3% LiCl aq. The solution was extracted withEtOAc twice and washed with brine. The organic layer was dried through(MgSO₄), and concentrated. The residue purified by CombiFlash on silicagel column using MeOH/DCM. To obtain 599 mg (64% yield) desiredtripeptide 86b. LCMS [M+H]⁺=489.88 (100%), Tr=1.05 min.

Compound 86

313 mg tripeptide 86b was converted to final compound 86 using asynthetic sequence identical to that reported in Example 84, yielding 7mg 86. ¹H NMR (400 MHz, Methanol-d₄): δ 8.80 (d, J=8.5 Hz, 1H), 8.22 (d,J=8.5 Hz, 1H), 7.88 (s, 1H), 7.80 (m, 1H), 7.55 (m, 1H), 7.42 (d, J=8.5Hz, 1H), 6.57-6.35 (dd, J=132.8, 16.3 Hz, 2H), 5.79 (q, J=7.1 Hz, 1H),5.28 (d, J=8.5 Hz, 1H), 5.05 (q, J=6.5 Hz, 1H), 4.39 (d, J=13.2 Hz, 1H),4.06 (m, 1H), 3.60 (dd, J=18.6, 5.4 Hz, 1H), 2.78 (s, 9H), 2.70-2.63 (m,1H), 2.27 (d, J=13.7 Hz, 1H), 1.90 (d, J=13.7 Hz, 1H), 1.68-1.63 (m,1H), 1.57 (dd, J=10.6, 7.0 Hz, 6H), 1.50 (s, 3H), 1.40 (s, 3H), 1.23 (m,12H). LCMS [M+H]⁺=608.25; Tr=1.17 min.

Compound 87

17 mg of 87 was the obtained as by-product during the synthesis of 86reported above. ¹H NMR (400 MHz, Methanol-d₄): δ 8.94 (d, J=8.5 Hz, 1H),8.22 (d, J=8.5 Hz, 1H), 7.81-7.77 (m, 2H), 7.58 (m, 1H), 7.40 (d, J=8.5Hz, 1H), 6.54-6.31 (dd, J=132.8, 16.3 Hz, 2H), 5.76 (q, J=7.1 Hz, 1H),5.33 (d, J=8.5 Hz, 1H), 5.05 (q, J=6.5 Hz, 1H), 4.40 (d, J=13.2 Hz, 1H),4.06 (m, 1H), 3.60 (dd, J=18.6, 5.4 Hz, 1H), 2.70-2.63 (m, 1H), 2.27 (d,J=13.7 Hz, 1H), 1.90 (d, J=13.7 Hz, 1H), 1.68-1.63 (m, 1H), 1.57 (dd,J=10.6, 7.0 Hz, 6H), 1.50 (s, 3H), 1.40 (s, 3H), 1.23 (m, 12H). LCMS[M+H]⁺=552.31; Tr=0.90 min.

Example 88 Compound 88

Compound 88a: 1,2-Dichloro-ethyne

Into an oven dried, argon purged flask were placed oil-free potassiumhydride (from 1740 mg of ca. 30% dispersion in mineral oil, ca. 13mmol), anhydrous tetrahydrofuran (10 mL) and hexane (1 mL). The flaskwas repurged with argon and trichloroethylene (900 μL, 1.32 g, 10 mmol)was added followed by dry methanol (10 μL, 7.9 mg, 0.25 mmol). Thismixture was stirred at room temperature for two hours. After this time,hexane (10 mL) was added and the resulting solution was immediately usedin the subsequent step.

Compound 88b: 4-Chloroethynyl-tetrahydro-pyran-4-carboxylic acid methylester

Into an oven dried, argon purged flask tetrahydrofuran (50 mL) was addedand the solution was cooled with an ice bath. A 1.8 M solution oflithium diisopropylamide (7.2 mL, 13 mmol) intetrahydrofuran/heptane/ethylbenzene was added. The resulting solutionwas cooled to −78° C., and treated dropwise with methyltetrahydro-2H-pyran-4-carboxylate (Sigma-Aldrich) (1.20 mL. 1.30 g, 9mmol) followed by hexamethylphosphoramide (1.56 mL, 1.61 g, 9 mmol). Theresulting solution was warmed to 0° C., stirred for 20 min., cooled to−78° C., and treated dropwise with pre-cooled (0° C.) solution of1,2-dichloro-ethyne (ca. 10 mmol). The reaction mixture was stirred at−78° C. for 30 min and then allowed to warm to room temperature. After 4hours at room temperature, the reaction mixture was poured into crushedice and extracted with diethyl ether (200 mL) (5 mL of brine was addedto support the separation). The organic phase was separated and washedwith water (200 mL). This water phase was extracted with diethyl ether(100 mL). The combined organic fractions were washed with brine (100mL), dried over magnesium sulfate, filtered through a 2 cm layer ofsilica gel (silica gel layer was washed with 50 mL of ethyl acetate),and then concentrated under reduced pressure. The crude product wassubjected to silica gel chromatography (gradient from 0-15% ethylacetate in iso-hexanes) to afford the title compound (1.22 g, 67%) as acolorless oil. R_(f)=0.48, 30% ethyl acetate in iso-hexanes,phosphomolybdic acid in ethanol. LCMS (m/z) 203.2/205.3 [M+H]′ Tr=3.12min (Gemini 5 u C18 110 Å, 50×4.60 mm 5 micron column, 3.5 min, 2ml/min, 5-100% acetonitrile/water, 0.1% acetic acid modifier gradient).¹H NMR (400 MHz, CDCl₃): δ3.79-3.65 (m, 4H), 3.72 (s, 3H), 1.97 (m, 2H),1.78 (m, 2H).

Compound 88c: 4-Ethynyl-tetrahydro-pyran-4-carboxylic acid methyl ester

4-Chloroethynyl-tetrahydro-pyran-4-carboxylic acid methyl ester (1.01 g,5 mmol) and copper powder (1.6 g, 25 mmol) were suspended intetrahydrofuran (100 mL). Acetic acid (15 mL) was added and the reactionmixture was heated to 60° C. for 3 hours. After this time, the reactionmixture was poured onto water (copper powder was filtered off with theuse of the filtration paper) and extracted with diethyl ether (3×50 mL).The combined organic extracts were washed with saturated solution ofammonium chloride (3×50 mL), with saturated solution of sodiumbicarbonate (2×50 mL) and with water (50 mL). This water phase wasextracted with diethyl ether (50 mL). Combined organic extracts werewashed with brine (50 mL) and dried over magnesium sulfate, filteredthrough a 2 cm layer of silica gel (silica gel layer was washed with 50mL of ethyl acetate), and concentrated under reduced pressure. Afterdrying under high vacuum for one day, the title compound was isolated(0.84 g, quantitative yield) as a colorless oil. R_(f)=0.37, 30% ethylacetate in iso-hexanes, phosphomolybdic acid in ethanol. LCMS (m/z)169.0 [M+H]′ Tr=2.43 min (Gemini 5 u C18 110 Å, 50×4.60 mm 5 microncolumn, 3.5 min, 2 ml/min, 5-100% acetonitrile/water, 0.1% acetic acidmodifier gradient). ¹H NMR (400 MHz, CDCl₃): δ3.80-3.67 (m, 4H), 3.73(s, 3H), 2.34 (s, 1H), 1.98 (m, 2H), 1.80 (m, 2H).

Compound 88d

To a mixture of methyl 4-vinyltetrahydro-2H-pyran-4-carboxylate(prepared following the hydrogenation of 88c with Lindlar's catalyst),(0.726 g, 4.27 mmol), triethylamine (1.29 g, 12.81 mmol) and the[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acid tert-butyl ester 39a(1.5 g, 4.27 mmol) in anhydrous dioxane (30 mL) were added palladiumacetate (0.19 g, 0.85 mmol) and tri-o-tolyphosphine (0.26 g, 0.85 mmol).The reaction mixture was heated to 100° C. for 2 hours and diluted withEtOAc (100 mL). The crude was washed with water and dried over Na₂SO₄.After concentration, the residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:0 to 0:1to afford 88a (1.1 g, 59%). LCMS (m/z) 441.2 [M+H]. Tr=2.21 min.

Compound 88e

To a solution of 88d (1.0 g, 2.27 mmol) in THF (10 mL) and MeOH (5 ml)was added 1 N KOH in water (11.4 mL). The reaction mixture was stirredat 50° C. for 2 hours and was acidified by adding 1 N HCl in water (11.4mL). After extracted with EtOAc (2×100 mL), dried over Na₂SO₄ andconcentrated, the crude acid was used without further purification. Thecrude acid above (0.328 g, 0.77 mmol), le (0.398, 0.92 mmol), 4-DMAP(0.225 g, 1.85 mmol), DIPEA (0.239 g, 1.85 mmol) and Shiina reagent(0.632 g, 1.84 mmol) were dissolved in DCM (10 mL). The reaction mixturewas stirred at room temperature for overnight. After concentration, thecrude was diluted with EtOAc (100 mL), washed with sat. NH₄Cl and driedover Na₂SO₄. After concentration, the residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:0 to 0:1to afford 88e (0.55 g, 85%). LCMS (m/z) 841.2 [M+H]. Tr=2.56 min.

Compound 88

To a solution of 88e (0.70 g, 0.83 mmol) in THF (10 mL) was added 1 NKOH in water (0.83 mL). The reaction mixture was vigorously stirred atroom temperature for 20 minutes and was acidified by adding 1 N HCl inwater (0.83 mL). After concentration, the crude was dissolved in 4 NHCl/dioxane (2 ml) and stirred for 1 hour. After concentration andco-evaporated with toluene (3×), the crude above was dissolved in DCM(800 mL). N,N-diisopropylethylamine (0.54 g, 4.2 mmol), DMAP (catalyticamount) and 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (0.63 g, 1.66 mmol). The reactionmixture was stirred at room temperature for 1 hour. After concentration,the crude was purified by prep-HPLC to obtain 88 (0.22 g, 45%) as awhite solid. ¹H NMR (400 MHz, Methanol-d₄): δ 8.81 (d, J=7.7 Hz, 1H),8.54-8.30 (m, 1H), 8.11-7.78 (m, 2H), 7.68 (dd, J=8.5, 1.7 Hz, 1H), 7.53(d, J=8.5 Hz, 1H), 6.62 (d, J=16.3 Hz, 1H), 6.34 (d, J=16.3 Hz, 1H),5.78 (m, 1H), 5.21 (dd, J=8.1, 1.0 Hz, 1H), 5.09 (q, J=6.7 Hz, 1H), 4.38(d, J=13.2 Hz, 1H), 3.92 (ddq, J=19.0, 12.0, 4.0, 3.4 Hz, 2H), 3.71-3.40(m, 3H), 2.69 (m, 1H), 2.38 (m, 1H), 2.17 (m, 3H), 2.08-1.74 (m, 4H),1.76-1.44 (m, 7H), 1.00 (dd, J=24.1, 6.7 Hz, 6H). LCMS (m/z) 592.3[M+H]. Tr=2.15 min.

Example 89 Compound 89

Compound 89a

To a mixture of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester 39a, (0.50 g, 1.42 mmol), triethylamine (0.43 g 4.26mmol) and the vinyl cyclohexanone ethoxy ester obtained from SmallMolecules, Inc. (0.279 g, 1.42 mmol) in anhydrous dioxane (15 mL) wereadded palladium acetate (0.063 g, 0.28 mmol) and tri-o-tolyphosphine(0.085 g, 0.28 mmol). The reaction mixture was heated to 100° C. for 3hours and diluted with EtOAc (50 mL). The crude mixture was washed withwater and dried over Na₂SO₄. After concentration, the residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 1:0 to 0:1 to afford 89a (0.30 g, 45%). LCMS(m/z) 467.2 [M+H]. Tr=2.39 min.

Compound 89b

To a solution of 89a (0.3 g, 0.64 mmol) in anhydrous THF (5 mL), cooledto −78° C., was added methyl magnesium bromide (3.0 M solution in ether,0.43 mL) dropwise. The mixture was stirred at −780° C. for 1 h. Thereaction mixture was diluted with EtOAc (30 mL) and quenched with sat.NH₄Cl (2 mL). After layers separation, the organic layer was dried overNa₂SO₄ and concentrated. the crude residue was purified by silica gelchromatography using iso-hexanes/ethyl acetate 1:0 to 1:1 to give thetile compound 89b (0.17 g, 56%) as a mixture of 1:1 cis/trans mixture.LCMS (m/z) 483.3 [M+H]. Tr=2.36 min.

Compound 89c

To a solution of 89b (0.17 g, 0.353 mmol) in THF (2 mL) and methanol (1mL) was added a solution of 1N KOH in water (1.76 mL). The reactionmixture was stirred at room temperature for 1 hour. 1 N HCl (1.76 mL)was added to the reaction mixture. After concentration, the crude wasunder high vacuum over night. The crude above and 2,6-lutidine (0.227 g,2.12 mmol) were dissolved in DCM (5 mL). The reaction mixture was cooledto 0° C., TBSOTf (0.280 g, 1.06 mmol) was added drop wise to thereaction mixture. After stirred at room temperature for 1 hour andconcentrated, the crude was purified by silica gel chromatography usingiso-hexanes/ethyl acetate 1:0 to 1:1 to give the 89c (0.150 g, 58%).LCMS (m/z) 741.4 [M+H]. Tr=3.56 min.

Compound 89d

A solution of 89c (0.15 g, 0.20 mmol) in tetrahydrofuran (1 mL) andmethanol (1 ml) was prepared and a 1M aq. solution of potassiumcarbonate (0.8 mL) was added. The reaction mixture was stirred at roomtemperature for 1 hour. Di-tert-butyl dicarbonate (0.44 g, 0.20 mmol)was added to the reaction solution. Then, the reaction mixture wasstirred at room temperature for an additional 1 h, and acidified to pH=4by adding 1 N HCl in water. After concentration and co-evaporated withtoluene (3×) to give the crude acid. The crude acid above, le (0.104 g,0.24 mmol), DMAP (0.06 g, 0.48 mmol), DIPEA (0.02 g, 0.48 mmol) andshiina reagent (0.138 g, 0.40 mmol) were dissolved in DCM (10 mL). Thereaction mixture was stirred at room temperature for overnight. Afterconcentration, the crude was diluted with EtOAc (100 mL), washed withsat. NH₄Cl and dried over Na₂SO₄. After concentration, the residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 1:0 to 0:1 to 89d (0.07 g, 35%). LCMS (m/z)983.5 [M+H]. Tr=2.98 min.

Compound 89

To a solution of 89d (0.06 g, 0.061 mmol) in THF (1 mL) was added 1 NKOH in water (0.061 mL). The reaction mixture was vigorously stirred atroom temperature for 20 minutes and was acidified by adding 1 N HCl inwater (0.1 mL). After concentration, the crude was dissolved in 4 NHCl/dioxane (3 ml) and stirred for 1 hour at room temperature. Afterconcentration and co-evaporated with toluene (3×), the crude above wasdissolved in DCM (20 mL). N,N-diisopropylethylamine (0.039 g, 0.31mmol), DMAP (catalytic amount) and2-(1H-7-azabenzotriazol-11-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (0.028 g, 0.073 mmol). The reactionmixture was stirred at room temperature for 1 hour. After concentration,the crude was purified by prep-HPLC to obtain 89 (0.006 g, 16%) as asingle compound. The other isomer was not pure. ¹H NMR (400 MHz,Methanol-d₄): δ 8.93 (d, J=8.6 Hz, 1H), 8.18 (d, J=8.6 Hz, 1H),8.14-8.02 (m, 2H), 7.86 (d, J=8.5 Hz, 1H), 7.04 (d, J=16.5 Hz, 1H), 6.81(d, J=16.5 Hz, 1H), 5.27 (dd, J=27.9, 7.0 Hz, 2H), 4.00 (s, 1H), 3.58(dd, J=9.0, 3.7 Hz, 1H), 3.06 (d, J=39.1 Hz, 1H), 2.32-1.77 (m, 11H),1.81-1.50 (m, 5H), 1.43 (s, 3H), 1.32 (d, J=6.9 Hz, 4H), 0.95 (d, J=6.9Hz, 3H), 0.80 (d, J=6.8 Hz, 3H). LCMS (m/z) 620.3 [M+H]. Tr=2.12 min.

Examples 90 and 91 Compound 90 and Compound 91

Compound 90a

To a solution of (R)-tert-butyl 1-(7-bromoquinolin-2-yl)ethylcarbamate39a (740 mg, 2.1 mmol) in anhydrous acetonitrile (25 mL) was addedpalladium(II) acetate (95 mg, 0.42 mmol), the indicated olefinicdimethylpyran ethoxycarboxylic ester (670 mg, 3.16 mmol),2-di-tert-butylphosphino-3,4,5,6-tetramethyl-2′,4′,6′-triisopropyl-1,1′-biphenyl(202 mg, 0.42 mmol) and triethylamine (630 mg, 6.3 mmol) then themixture was heated in the microwave at 150° C. for 2 hours. The reactionmixture was filtered through Celite and the filter pad was washed withtetrahydrofuran (20 mL). The filtrate was evaporated then water (50 mL)was added and the organics extracted with ethyl acetate (2×50 mL). Thecombined organics were dried over sodium sulfate, filtered andconcentrated under reduced pressure to give a crude residue. This wassubjected to silica gel chromatography (gradient from 0-70% ethylacetate in iso-hexanes) to afford 90a (170 mg, 17%) as a white solid. ¹HNMR (400 MHz, CDCl₃): δ 8.04 (d, J=8.4 Hz, 1H), 7.96 (s, 1H), 7.71 (d,J=8.5 Hz, 1H), 7.53 (d, J=8.5 Hz, 1H), 7.27 (d, J=8.4 Hz, 1H), 6.60 (d,J=16.3 Hz, 1H), 6.35 (d, J=16.3 Hz, 1H), 6.20-6.10 (m, 1H), 5.01-4.89(m, 1H), 4.25-4.09 (m, 2H), 3.91-3.69 (m, 2H), 2.41-2.14 (m, 2H),1.80-1.61 (m, 2H), 1.56 (s, 3H), 1.45 (s, 9H), 1.32-1.21 (m, 6H), 1.15(s, 3H). LCMS (m/z) 483.2 [M+H], Tr=4.28 min.

Compound 90b

A solution of 90a (160 mg, 0.33 mmol) in tetrahydrofuran/water/methanol(10 mL, 4:1:1) was treated with lithium hydroxide (12 mg, 0.50 mmol).After stirring at 60° C. for 6 hours, the reaction was concentrated downunder reduced pressure. The residue was dissolved in water (20 mL), 1 Mhydrochloric acid (0.5 mL, 0.5 mmol) was added and the mixture wasquickly extracted with dichloromethane (2×50 mL). The organics werecombined, washed with brine, dried over sodium sulfate and filtered. Thefiltrate was evaporated to afford 90b (110 mg, 73%) as a white solid. ¹HNMR (400 MHz, CDCl₃): δ 8.03 (d, J=8.2 Hz, 1H), 7.96 (s, 1H), 7.70 (d,J=8.5 Hz, 1H), 7.58 (d, J=8.5 Hz, 1H), 7.22 (d, J=8.2 Hz, 1H), 6.62 (d,J=16.3 Hz, 1H), 6.35 (d, J=16.3 Hz, 1H), 6.17-6.15 (m, 1H), 5.21-4.94(m, 1H), 3.92-3.72 (m, 2H), 2.43-2.15 (m, 2H), 1.82-1.63 (m, 2H), 1.61(s, 3H), 1.42 (s, 9H), 1.24 (s, 3H), 1.15 (s, 3H). LCMS (m/z) 455.2[M+H], Tr=3.35 min (Gemini 5 u C18 110 Å, 50×4.60 mm 5 micron column, 6min, 2 ml/min, 5-100% acetonitrile/water, 0.1% acetic acid modifiergradient).

Compound 90c

Into an oven-dried, argon purged flask were placed2-methyl-6-nitrobenzoic anhydride (125 mg, 0.36 mmol),4-dimethylaminopyridine (62 mg, 0.51 mmol), 90b (110 mg, 0.24 mmol), andanhydrous dichloromethane (10 mL). Into the resulting solution was addedN,N-diisopropylethylamine (66 mg, 0.51 mmol) and this reaction mixturewas stirred at room temperature for 10 minutes. 1e (157 mg, 0.36 mmol)was added dropwise via syringe as a solution in anhydrousdichloromethane (5 mL). After stirring for 12 hours at room temperature,the reaction mixture was transferred to a separatory funnel and washedwith water (20 mL, 10 mL of brine was added to support the separation).The aqueous phase was extracted with dichloromethane (20 mL). Combinedorganic extracts were washed with brine (20 mL) and dried over magnesiumsulfate, filtered and concentrated under reduced pressure. The residuewas purified by silica gel chromatography (gradient from 0-40% ethylacetate+methanol (4/1) in iso-hexanes) to afford 90c (201 mg, 96%) as awhite solid after evaporation. ¹H NMR (400 MHz, CD₃OD): δ 8.28 (d, J=8.4Hz, 1H), 7.98 (s, 1H), 7.88 (d, J=8.2 Hz, 1H), 7.78 (d, J=9.4 Hz, 1H),7.53 (d, J=8.4 Hz, 1H), 6.93-6.81 (m, 1H), 6.67-6.51 (m, 1H), 5.51-5.34(m, 2H), 5.03 (d, J=12.1 Hz, 2H), 4.86 (d, J=12.1 Hz, 2H), 4.13 (q,J=7.1 Hz, 1H), 3.91-3.83 (m, 3H), 2.49-2.33 (m, 2H), 2.30-2.20 (m, 1H),2.17-2.07 (m, 3H), 2.02-1.85 (m, 2H), 1.83-1.65 (m, 2H), 1.53 (d, J=7.1Hz, 3H), 1.47 (s, 9H), 1.34 (d, J=6.9 Hz, 3H), 1.04 (d, J=6.9 Hz, 6H),0.88 (d, J=6.8 Hz, 6H). LCMS (m/z) 868.2/870.2 [M+H]′ Tr=3.22 min(Gemini 5 u C18 110 Å, 50×4.60 mm 5 micron column, 3.5 min, 2 ml/min,5-100% acetonitrile/water, 0.1% acetic acid modifier gradient).

Compounds 90 and 91

To 90c (195 mg, 0.22 mmol) in tetrahydrofuran (10 mL) was added asolution of lithium hydroxide (6 mg, 0.53 mmol) in water (5 mL). Afterstirring at room temperature for 2 hour, 1M hydrochloric acid was added(0.25 mL of 1M solution in water, 0.25 mmol) and the reaction mixturewas concentrated under reduced pressure and the residue partitionedbetween dichloromethane (50 mL) and water (50 mL). The organic layer wascollected, concentrated under reduced pressure and dried under highvacuum for one day. The residue was treated with 4M hydrogen chloride in1,4-dioxane (5 mL, 20 mmol) at room temperature under argon for 4 hours.Reaction mixture was concentrated under reduced pressure. This residuewas dissolved in N,N-dimethylformamide (5 mL) and the obtained solutionwas added into an argon purged flask containing2-(1H-7-azabenzotriazol-1-yl)-11,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (128 mg, 0.34 mmol),N,N-diisopropylethylamine (144 mg, 1.12 mmol) and dichloromethane (100mL). The reaction mixture was re-purged with argon and stirred at roomtemperature for 2 hours. The obtained reaction mixture was washed withwater (100 ml) and brine (100 mL), dried over magnesium sulfate,filtered and concentrated under reduced pressure. The residue waspurified by silica gel chromatography (gradient from 0-40% ethyl acetatein iso-hexanes) to afford firstly Compound 90 (27 mg, 19%) and thenCompound 91 (17 mg, 12%) white solids after evaporation.

Compound 90: ¹H NMR (400 MHz, CD₃OD): δ 9.08 (d, J=8.6 Hz, 1H), 8.48 (s,1H), 8.27 (d, J=8.6 Hz, 1H), 8.03 (s, 1H), 7.97 (dd, J=8.5, 1.5 Hz, 1H),6.91-6.69 (m, 2H), 5.92-5.73 (m, 1H), 5.36-5.22 (m, 1H), 4.42-4.26 (m,1H), 4.01-3.83 (m, 2H), 3.83-3.73 (m, 1H), 3.71-3.62 (m, 1H), 3.55-3.42(m, 1H), 2.53-2.41 (m, 1H), 2.33-2.26 (m, 1H), 2.13-2.05 (m, 1H),2.00-1.87 (m, 2H), 1.81 (d, J=7.2 Hz, 3H), 1.79-1.61 (m, 4H), 1.44-1.38(m, 2H), 1.33 (s, 3H), 1.30-1.24 (m, 3H), 1.22 (s, 3H), 1.11-1.02 (m,3H), 0.99 (d, J=6.8 Hz, 3H). LCMS (m/z) 620.4 [M+H]′ Tr=3.45 min

Compound 91: ¹H NMR (400 MHz, CD₃OD): δ8.27 (d, J=8.5 Hz, 1H), 7.86 (d,J=8.5 Hz, 1H), 7.83 (s, 1H), 7.64 (dd, J=8.4, 1.7 Hz, 1H), 7.47 (d,J=8.5 Hz, 1H), 6.63 (d, J=16.2 Hz, 1H), 6.24 (d, J=16.3 Hz, 1H), 5.82(q, J=7.2 Hz, 1H), 5.24 (d, J=8.3 Hz, 1H), 5.16-5.05 (m, 1H), 4.53-4.40(m, 1H), 3.91-3.82 (m, 2H), 3.67-3.57 (m, 1H), 2.72 (td, J=12.9, 3.3 Hz,1H), 2.54-2.44 (m, 1H), 2.40 (dd, J=13.8, 1.9 Hz, 1H), 2.34-2.16 (m,2H), 2.02-1.92 (m, 1H), 1.90-1.79 (m, 1H), 1.78-1.70 (m, 2H), 1.68-1.52(m, 2H), 1.64 (d, J=7.2 Hz, 3H), 1.61 (d, J=6.7 Hz, 3H), 1.31 (s, 3H),1.20 (s, 3H), 1.12 (d, J=6.8 Hz, 3H), 1.08 (d, J=6.6 Hz, 3H). LCMS (m/z)620.4 [M+H]′ Tr=3.47 min (Gemini 5 u C18 110 Å, 50×4.60 mm 5 microncolumn, 6 min, 2 ml/min, 5-100% acetonitrile/water, 0.1% acetic acidmodifier gradient).

Example 92 Compound 92

Compound 92a

The commercially supplied ethyl-4-oxo-1-vinylcyclohexanecarboxylate(provided by Small Molecules, Inc.), (3.0 g, 15.29 mmol) in MeOH (80 mL)in a flask equipped with a dropping funnel was added a MeOH (80 mL)solution of NaBH₄ (697 mg, 18.35 mmol) dropwise at 0° C. It was stirredat 0° C. for 10 min, and RT for 48 h. After quenched with sat'd NH₄Clsolution, it was extracted with EtOAc and washed with brine once. Theorganic layer was dried through (Na₂SO₄), and concentrated. The cis andtrans isomers were separated by CombiFlash on silica gel column usingEtOAC/Hexane as eluents. On TLC with EA/HEX=1/3, Rf of trans=0.5, Rf ofcis=0.4. Yield: trans-isomer 92a was 425 mg, cis-isomer was 864 mg, mixof cis/trans was 468 mg. Total yield of (cis+trans) was 1.76 g, 59%.

For trans-isomer 92a: ¹H NMR (400 MHz, Chloroform-d): δ 5.84-5.71 (m,1H), 5.23-5.11 (m, 2H), 4.11 (q, J=7.5 Hz, 2H), 3.79 (tq, J=6.8, 3.6 Hz,1H), 1.90 (ddd, J=11.7, 8.2, 4.2 Hz, 4H), 1.71 (ddt, J=11.9, 7.8, 3.7Hz, 2H), 1.61-1.50 (m, 2H), 1.22 (t, J=7.1 Hz, 3H).

For cis-isomer: ¹H NMR (400 MHz, Chloroform-d): δ 5.78 (dd, J=17.5, 10.6Hz, 1H), 5.13-5.00 (m, 2H), 4.15 (q, J=7.1 Hz, 2H), 3.61 (dt, J=13.2,6.4 Hz, 1H), 2.38-2.22 (m, 2H), 1.89 (dt, J=10.7, 5.2 Hz, 2H), 1.47-1.29(m, 4H), 1.24 (t, J=7.1 Hz, 3H).

Compound 92b

The reactant 92a (1R,4R)-ethyl 4-hydroxy-1-vinylcyclohexanecarboxylate(425 mg, 2.13 mmol) in DCM (10 mL) was added with CH₃OBF₄ (1.66 g, 10.7mmol) and Proton Spronge (2.3 g, 10.7 mmol) and stirred at RT for 24 h.After filtrated through a pile of Celite, the filtrate was washed withsat'd NaHCO₃ solution. The organic layer was dried through (Na₂SO₄), andconcentrated and purified by CombiFlash on silica gel column usingEtOAC/Hexane as eluents. It gave 223 mg (49% yield) of (1R,4R)-ethyl4-methoxy-1-vinyl-cyclohexanecarboxylate 92b. ¹H NMR (400 MHz,Chloroform-d): δ 5.87-5.68 (m, 1H), 5.20-4.99 (m, 2H), 4.23-3.95 (m,2H), 3.36-3.19 (m, 4H), 1.87 (pd, J=13.5, 5.6 Hz, 4H), 1.75-1.54 (m,4H), 1.29-1.14 (m, 3H).

Compound 92c

Proceeding from 223 mg (1.05 mmol) 92b, Heck reaction to form(1R,4R)-ethyl1-((E)-2-(7-((R)-1-(tert-butoxycarbonylamino)ethyl)naphthalen-2-yl)vinyl)-4-methoxycyclohexanecarboxylatewas done in the same manner as in Example 74, instead using ethyl4-oxo-1-vinylcyclohexanecarboxylate, to give 424 mg (89% yield)intermediate compound 92c. LCMS [M+H]⁺=455.10

Compound 92

Compound 92 was prepared using a multistep synthesis beginning from 424mg (0.92 mmol) 92c where esterification with 1e, selective hydrolysisand then N-Boc removal provided an intermediate which furnishedmacrocycle 92 (36 mg) in 6.2% overall yield after treatment withHATU/DIPEA in DCM/DMF using the lactonization and final purificationmethod reported in Example 90. ¹H NMR (400 MHz, Methanol-d₄): δ 8.21 (d,J=8.5 Hz, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.75 (d, J=1.7 Hz, 1H), 7.59 (dd,J=8.5, 1.7 Hz, 1H), 7.41 (d, J=8.5 Hz, 1H), 6.40 (dd, J=124.5, 16.3 Hz,2H), 5.78 (q, J=7.2 Hz, 1H), 5.23 (d, J=8.6 Hz, 1H), 5.09-4.99 (m, 1H),4.45-4.34 (m, 1H), 3.60-3.51 (m, 1H), 3.44 (s, 1H), 3.33 (s, 3H), 2.66(td, J=13.0, 3.3 Hz, 1H), 2.29-2.09 (m, 3H), 2.05-1.78 (m, 6H),1.74-1.62 (m, 2H), 1.62-1.39 (m, 8H), 1.26 (m, 3H), 1.02 (dd, J=19.3,6.7 Hz, 6H). LCMS [M+H]⁺=620.63.

Example 93 Compound 93

Compound 93a. (S)-4-Benzyl-3-(2-cyclopropyl-acetyl)-oxazolidin-2-one

A cooled (−40° C.) solution of cyclopropylacetic acid (5.13 g, 51.250mmol) in anhydrous tetrahydrofuran (150 mL) was subsequently treatedwith triethylamine (21.4 mL, 153.75 mmol) and trimethylacetyl chloride(6.9 mL, 56.375 mmol). After stirring at −40° C. for 1 h, the mixturewas treated with (S)-4-benzyl-oxazolidin-2-one (9.99 g, 56.375 mmol) andlithium chloride (2.39 g, 56.375 mmol). The mixture was allowed toslowly warm to room temperature and after stirring at room temperaturefor 22 h, the reaction was partitioned between dichloromethane andhalf-saturated ammonium chloride. The aqueous layer was extracted withdichloromethane. The organics were combined, filtered through a phaseseparator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 100 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to9:1 to afford the title compound (11.63 g, 87%) as a colorless oil. ¹HNMR (300 MHz, CDCl₃) δ0.19-0.32 (m, 2H), 0.56-0.69 (m, 2H), 1.10-1.24(m, 1H), 2.76-2.87 (m, 2H), 2.96 (dd, J=16.9, 6.9 Hz, 1H), 3.34 (dd,J=13.2, 3.3 Hz, 1H), 4.15-4.29 (m, 2H), 4.66-4.77 (m, 1H), 7.20-7.40 (m,5H). LCMS (m/z) 282.1 [M+Na], Tr=2.78 min.

Compound 93b.(S)-4-Benzyl-3-((S)-2-cyclopropyl-2-hydroxy-acetyl)-oxazolidin-2-one

A cooled (−78° C.) solution of sodium bis(trimethylsilyl)amide (0.6 M intoluene, 65 mL, 38.918 mmol) was diluted with anhydrous tetrahydrofuran(100 mL) and treated with a solution of(S)-4-benzyl-3-(2-cyclopropyl-acetyl)-oxazolidin-2-one (9.174 g, 35.380mmol) in anhydrous tetrahydrofuran (50 mL). After stirring at −78° C.for 45 min, the mixture was treated with a solution of2-benzenesulfonyl-3-phenyl-oxaziridine (11.064 g, 42.456 mmol) inanhydrous tetrahydrofuran (50 mL). After stirring at −78° C. for 55 min,the reaction was quenched with acetic acid (10.1 mL, 176.9 mmol) andallowed to warm to room temperature. The mixture was quenched withsodium bicarbonate solution (300 mL) and poured overdichloromethaneliso-hexanes (3:1, 400 mL). The aqueous layer wasextracted with dichloromethane and the organics were combined, washedwith saturated sodium thiosulfate, hydrochloric acid (1 M, 300 mL),water and filtered through a phase separator. The volatiles were removedin vacuo and the residue was purified by silica gel chromatography usinga 100 g Isolute cartridge eluted with a continuous gradient ofiso-hexanes/ethyl acetate 1:0 to 7:3 followed by silica gelchromatography using a 100 g Isolute cartridge eluted with a continuousgradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the titlecompound (7.922 g, 81%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃)δ0.42-0.65 (m, 4H), 1.25-1.38 (m, 1H), 2.88 (dd, J=13.4, 9.6 Hz, 1H),3.35 (dd, J=13.4, 3.4 Hz, 1H), 3.44 (d, J=8.2 Hz, 1H), 4.25-4.37 (m,2H), 4.67-4.72 (m, 1H), 4.80 (dd, J=8.0, 6.0 Hz, 1H), 7.21-7.41 (m, 5H).LCMS (m/z) 298.1 [M+Na], Tr=2.27 min.

Compound 93c.(S)-4-Benzyl-3-((S)-2-cyclopropyl-2-triisopropylsilanyloxy-acetyl)-oxazolidin-2-one

A cooled (0° C.) solution of(S)-4-benzyl-3-((S)-2-cyclopropyl-2-hydroxy-acetyl)-oxazolidin-2-one(4.137 g, 15.026 mmol) in dichloromethane (60 mL) was subsequentlytreated with 2,6-lutidine (3.5 mL, 30.052 mmol) and triisopropylsilyltrifluoromethanesulfonate (6.1 mL, 22.539 mmol). After stirring at roomtemperature for 1 h, the reaction was quenched with pH 4 buffer(phthalate). The aqueous layer was extracted with dichloromethane. Theorganics were combined, filtered through a phase separator and thevolatiles were removed in vacuo. The residue was purified by silica gelchromatography using a 100 g Isolute cartridge eluted with a continuousgradient of iso-hexanes/diethyl ether 1:0 to 9:1 to afford the titlecompound (5.28 g, 81%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃)δ0.40-0.49 (m, 2H), 0.53-0.62 (m, 1H), 0.63-0.71 (m, 1H), 1.02-1.18 (m,21H), 1.20-1.34 (m, 1H), 2.66 (dd, J=13.1, 10.5 Hz, 1H), 3.49 (dd,J=13.1, 3.4 Hz, 1H), 4.17-4.27 (m, 2H), 4.63-4.73 (m, 1H), 5.58 (d,J=5.1 Hz, 1H), 7.24-7.40 (m, 5H).

Compound 93d. (S)-Cyclopropyl-hydroxy-acetic acid

A cooled (0° C.) solution of(S)-4-benzyl-3-((S)-2-cyclopropyl-2-triisopropylsilanyloxy-acetyl)-oxazolidin-2-one(2.260 g, 5.235 mmol) in tetrahydrofuran/water (60 mL, 5:1) wassubsequently treated with hydrogen peroxide (30% aqueous, 2.7 mL, 26.173mmol) and lithium hydroxide monohydrate (439.2 mg, 10.470 mmol). Afterstirring at 0° C. for 4 h, the reaction was quenched with concentratedhydrochloric acid (˜8 mL) and sodium metabisulfite (9.9 g). The aqueouslayer was extracted with tetrahydrofuran/ethyl acetate (2×). Theorganics were combined, filtered through a phase separator and thevolatiles were removed in vacuo. The residue was purified by silica gelchromatography using a 25 g Isolute cartridge eluted with a continuousgradient of dichloromethane/methanol 1:0 to 9:1 to afford the titlecompound (298.3 mg, 49%) as colorless needles. ¹H NMR (300 MHz, CDCl₃)δ0.45-0.71 (m, 4H), 1.14-1.29 (m, 1H), 3.87 (d, J=6.9 Hz, 1H).

Compound 93e.((S)-2-{(S)-3-[(R)-1-(7-Bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

A cooled (0° C.) solution of(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (3.007 g, 6.948 mmol) intetrahydrofuran/water (60 mL, 5:1) was treated with lithium hydroxidemonohydrate (874.4 mg, 20.844 mmol). After stirring at 0° C. for 40minutes the reaction was quenched with hydrochloric acid (1 M, 50 mL).The aqueous layer was extracted with ethyl acetate (2×). The organicswere combined, dried over anhydrous sodium sulfate, filtered and thevolatiles were removed in vacuo. Residual trichlorethanol was azeotropedoff with toluene (3×) to provide(S)-1-((S)-2-tert-butoxycarbonylamino-propionyl)-hexahydro-pyridazine-3-carboxylicacid as a white solid which was then combined with(R)-1-(7-bromo-quinolin-2-yl)-ethylamine hydrochloride (1.998 g, 6.948mmol) and suspended in anhydrous acetonitrile (60 mL) andtetrahydrofuran (10 mL). The suspension was cooled to 0° C. andsubsequently treated with N,N-diisopropylethylamine (6 mL, 34.740 mmol)and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (3.699 g, 9.727 mmol). After slowly warmingto room temperature and stirring for 16 h, the reaction was quenched at0° C. with hydrochloric acid (1 M, 70 mL). The aqueous layer wasextracted with ethyl acetate (2×). The organics were combined, washedwith a saturated solution of sodium bicarbonate, dried over anhydroussodium sulfate, filtered and the volatiles were removed in vacuo. Theresidue was purified by silica gel chromatography using a 100 g Isolutecartridge eluted with a continuous gradient of iso-hexanes/ethyl acetate1:0 to 1:4 to afford the title compound (3.702 g, 99%) as a white foam.¹H NMR (300 MHz, CDCl₃) δ1.38 (d, J=6.9 Hz, 3H), 1.43 (s, 9H), 1.47-1.55(m, 2H), 1.58 (d, J=6.9 Hz, 3H), 1.60-1.78 (m, 2H), 2.22-2.31 (m, 1H),2.65-2.78 (m, 1H), 3.39-3.52 (m, 1H), 4.55 (d, J=13.4 Hz, 1H), 5.18-5.34(m, 2H), 5.36-5.45 (m, 1H), 7.37 (d, J=8.5 Hz, 1H), 7.65 (dd, J=8.5, 1.6Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 8.04 (d, J=6.5 Hz, 1H), 8.14 (d, J=8.5Hz, 1H), 8.36 (s, 1H). LCMS (m/z) 536.1, 537.1 [M+H], Tr=2.58 min.

Compound 93f.(S)-1-[(S)-2-((S)-2-Cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

A cooled (0° C.) solution of(S)-2-{(S)-3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester (802.2 mg, 1.501 mmol) in dichloromethane (20 mL)was treated with a solution of hydrogen chloride (1.9 mL, 7.505 mmol, 4M in 1,4-dioxane). After stirring at room temperature for 3 h, thevolatiles were removed in vacuo. Residual water was azeotroped off withtoluene to provide(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide hydrochloride as a whitesolid which was then combined with (S)-cyclopropyll-hydroxy-acetic acid(174.3 mg, 1.501 mmol) and dry acetonitrile (20 mL). This suspension wascooled to 0° C. and subsequently treated with N,N-diisopropylethylamine(0.79 mL, 4.503 mmol) and(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(796.6 mg, 1.801 mmol). The reaction was slowly warmed to roomtemperature. After stirring for 16 h the reaction was quenched at 0° C.with hydrochloric acid (1 M, 40 mL) and diluted with dichloromethane.The aqueous layer was extracted with dichloromethane. The organics werewashed with a saturated solution of sodium bicarbonate. The basicaqueous layer was extracted with dichloromethane. The organics werecombined, filtered through a phase separator and the volatiles wereremoved in vacuo. The residue was purified by silica gel chromatographyusing a 50 g Isolute cartridge eluted with a continuous gradient ofdichloromethane/methanol 1:0 to 95:5 to afford the title compound (393.0mg, 49%) as a white solid. ¹H NMR (300 MHz, CDCl₃) δ 0.43-0.72 (m, 4H),1.03-1.15 (m, 1H), 1.47 (d, J=6.7 Hz, 3H), 1.59 (d, J=6.7 Hz, 3H),1.64-1.78 (m, 2H), 1.93-2.03 (m, 1H), 2.23-2.33 (m, 1H), 2.65-2.77 (m,1H), 3.44-3.58 (m, 2H), 3.62-3.75 (m, 2H), 3.80 (d, J=11.8 Hz, 1H),4.49-4.59 (m, 1H), 5.28 (app pentet, J=6.7 Hz, 1H), 5.49 (app pentet,J=7.8 Hz, 1H), 7.20 (d, J=8.2 Hz, 1H), 7.40 (d, J=8.5 Hz, 1H), 7.64 (dd,J=8.7, 1.8 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 8.15 (d, J=8.5 Hz, 1H), 8.48(s, 1H). LCMS (m/z) 534.0 [M+H], Tr=2.07 min.

Compound 93g.6-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-[1,4]dioxepane-6-carboxylicacid

A solution of(S)-1-[(S)-2-((S)-2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (244 mg, 0.459 mmol),6-vinyl-[1,4]dioxepane-6-carboxylic acid 57d (79 mg, 0.459 mmol),triethylamine (140 mg, 192 μL, 1.38 mmol) and tri(o-tolyl)phosphine (28mg, 0.092 mmol) in 1,4-dioxane (9 mL) was degassed with nitrogen for 5min and then warmed to 50° C. under nitrogen with stirring.Tris(dibenzylideneacetone)dipalladium(0) (42 mg, 0.046 mmol) was addedand the mixture was heated to 80° C. for 45 min and then allowed tocool. The suspension was filtered and evaporated to give crude titlecompound which was used in the next stage without further purification.LCMS (m/z) 624.2 [M+H], Tr=1.58 min.

Compound 93

To a stirred mixture of powdered 4 Å molecular sieves (˜1 g),2-methyl-6-nitrobenzoic anhydride (316 mg, 0.918 mmol) and4-(dimethylamino)-pyridine (224 mg, 1.84 mmol) in dichloromethane (138mL), under nitrogen, was added a solution of 93g in dichloromethane (10mL) over 3 h via syringe pump. The flask originally containing the crudeacid was washed with dichloromethane (1 mL) and this was added to thereaction mixture over ca. 20 min. After the end of the addition, thereaction mixture was stirred for 1.5 h, filtered through Celite andwashed successively with ice-cold saturated ammonium chloride solution,ice-cold saturated sodium bicarbonate solution, saturated brine andfiltered through a hydrophobic frit and evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/acetone 1:1 to 1:3 followed by reverse phase preparativeHPLC using a gradient of acetonitrile/water 1:9 to 7:3 to give, aftertrituration with diethyl ether, the title compound (21 mg, 8% over 2steps) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ0.49-0.74 (m, 4H),1.27-1.41 (m, 1H), 1.48-1.78 (m, 2H), 1.60 (d, J=6.9 Hz, 3H), 1.65 (d,J=7.3 Hz, 3H), 1.91-2.01 (m, 1H), 2.20-2.29 (m, 2H), 2.65-2.76 (m, 1H),3.57-3.94 (m, 5H), 4.17 (d, J=12.7 Hz, 1H), 4.36 (d, J=12.7 Hz, 1H),4.39-4.48 (m, 1H), 4.53 (d, J=12.7 Hz, 1H), 5.03-5.12 (m, 3H), 5.84 (q,J=7.1 Hz, 1H), 6.33 (d, J=16.5 Hz, 1H), 6.52 (d, J=16.5 Hz, 1H), 7.47(d, J=8.5 Hz, 1H), 7.70 (dd, J=8.5, 1.6 Hz, 1H), 7.78 (br s, 1H), 7.86(d, J=8.5 Hz, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 606.2 [M+H],Tr=2.08 min.

Example 94 Compound 94

Compound 94a. 3-Methyl-tetrahydro-pyridazine-1,2,3-tricarboxylic acid1,2-di-tert-butyl ester 3-methyl ester

To a solution of (S)-tetrahydro-pyridazine-1,2,3-tricarboxylic acid1,2-di-tert-butyl ester 3-(2,2,2-trichloro-ethyl) ester (5.0 g, 10.8mmol) in tetrahydrofuran/methanol (50 mL, 1:1) containing 3 Å molecularsieves at 0° C. was added tetra n-butylammonium fluoride (22 mL, 22mmol, 1 M in tetrahydrofuran). After stirring at room temperature for 24h, the mixture was filtered through a pad of Celite and the filtrate wasevaporated. The residue was partitioned between diethyl ether andsaturated sodium hydrogen carbonate solution. The aqueous layer wasextracted with diethyl ether. The organic extracts were combined and thesolution was filtered through a hydrophobic frit and the solvent wasevaporated. The residue was purified by silica gel chromatographyeluting with iso-hexanes/ethyl acetate 20:1 to 3:2 to afford(S)-tetrahydro-pyridazine-1,2,3-tricarboxylic acid 1,2-di-tert-butylester 3-methyl ester (3.0 g, 82%) as a colorless oil and as a mixture ofrotamers. LCMS (m/z) 345.1 [M+H], Tr=2.78 min. A solution of(S)-tetrahydro-pyridazine-1,2,3-tricarboxylic acid 1,2-di-tert-butylester 3-methyl ester (3.0 g, 8.72 mmol) in tetrahydrofuran (20 mL) at−78° C. was treated with lithium bis(trimethylsilyl)amide (13 mL, 13.0mmol, 1 M in tetrahydrofuran). After stirring at −78° C. for 1 h, themixture was treated with iodomethane (1.4 mL, 22.5 mmol). The reactionmixture was stirred at −78° C. for 1 h, then at 0° C. for 1 h and atroom temperature for 1 h. The reaction mixture was re-cooled to 0° C.and quenched with water. The aqueous phase was extracted withdichloromethane (2×) and the organic solution was passed through ahydrophobic frit and the solvent was evaporated. The residue waspurified by silica gel chromatography eluting with a gradient ofiso-hexanes/ethyl acetate 4:1 to 0:1 to afford the title compound (1.62g, 52%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ1.41-1.52 (m, 18H),1.53-1.67 (m, 4H), 1.71-1.99 (m, 3H), 3.17-3.51 (m, 1H), 3.70-3.81 (m,3H), 3.90-4.21 (m, 1H). LCMS (m/z) 359.0 [M+H], Tr=2.90 min.

Compound 94b.3-[(R)-1-(7-Bromo-quinolin-2-yl)-ethylcarbamoyl]-3-methyl-tetrahydro-pyridazine-1,2-dicarboxylicacid di-tert-butyl ester

To a solution of 3-methyl-tetrahydro-pyridazine-1,2,3-tricarboxylic acid1,2-di-tert-butyl ester 3-methyl ester (1.62 g, 4.52 mmol) intetrahydrofuran/water/methanol (50 mL, 5:1:1) at 0° C. was added lithiumhydroxide hydrate (2.2 g, 52.3 mmol). The reaction mixture was stirredat 0° C. for 1 h and then at room temperature for 1 h. The reactionmixture was heated at 40° C. for 19 h then cooled to room temperatureand the solvent was evaporated. The residue was partitioned betweenethyl acetate and water and the mixture was acidified to pH 5 withhydrochloric acid (1 M). The organic phase was separated and the aqueousphase was extracted with ethyl acetate. The combined organic extractswere dried over anhydrous magnesium sulfate, filtered and the solventwas evaporated. The residue was dissolved in toluene and the solvent wasevaporated to give 3-methyl-tetrahydro-pyridazine-1,2,3-tricarboxylicacid 1,2-di-tert-butyl ester as a yellow oil (1.47 g, 95%). LCMS (m/z)343.1 [M−H], Tr=2.36 min. To a solution of3-methyl-tetrahydro-pyridazine-1,2,3-tricarboxylic acid1,2-di-tert-butyl ester (1.37 g, 4.0 mmol) in dichloromethane (30 mL) at0° C. was added (R)-1-(7-bromo-quinolin-2-yl)-ethylamine hydrochloride(1.30 g, 4.5 mmol) and N,N-diisopropylethylamine (2.8 mL, 16.0 mmol)followed by(dimethylamino)-N,N-dimethyl(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methaniminiumhexafluorophosphate (1.81 g, 4.76 mmol). The reaction mixture wasstirred at 0° C. for 30 min and then at room temperature for 2 h. Thereaction mixture was diluted with dichloromethane and washed with water.The layers were separated and the aqueous layer was extracted withdichloromethane. The combined organic extracts were passed through ahydrophobic frit and the solvent was evaporated. The residue waspurified by silica gel chromatography eluting with a gradient ofiso-hexanes/ethyl acetate 20:1 to 0:1 to afford the title compound (929mg, 40%) as a yellow foam and as a mixture of diastereoisomers. LCMS(m/z) 577.3, 579.2 [M+H], Tr=3.72 min.

Compound 94c.((S)-2-{3-[(R)-1-(7-Bromo-quinolin-2-yl)-ethylcarbamoyl]-3-methyl-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester

To a solution of3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-3-methyl-tetrahydro-pyridazine-1,2-dicarboxylicacid di-tert-butyl ester (925 mg, 1.6 mmol) in dichloromethane (10 mL)at 0° C. was added trifluoroacetic acid (3 mL). The reaction mixture wasstirred at 0° C. for 30 min and then at room temperature for 36 h. Thereaction mixture was concentrated at reduced pressure and the residuewas co-evaporated with toluene (3×) to give3-methyl-hexahydro-pyridazine-3-carboxylic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide bis-trifluoroacetate salt(1.6 mmol) as a light brown foam. LCMS (m/z) 377.3, 379.2 [M+H], Tr=1.18min. To a solution of (S)-2-tert-butoxycarbonylamino-propionic acid (303mg, 1.6 mmol) in anhydrous acetonitrile (12 mL) at 0° C. was addedO-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(915 mg, 2.41 mmol) and N,N-diisopropylethylamine (1.5 mL, 8.42 mmol).The reaction mixture was stirred at 0° C. for 15 min and then treatedwith a solution of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amidebis-trifluoroacetate salt (1.6 mmol) in anhydrous acetonitrile (8 mL).The reaction mixture was allowed to warm to room temperature and wasstirred for 16 h. The solvent was evaporated and the residue waspartitioned between dichloromethane and water. The organic layer wasseparated and the aqueous layer was extracted with dichloromethane. Thecombined organic extracts were passed through a hydrophobic frit and thesolvent was evaporated. The residue was purified by silica gelchromatography eluting with a gradient of iso-hexanes/ethyl acetate 20:1to 0:1 to afford the title compound (857 mg, 40% over 2 steps) as abrown oil and as a mixture of diastereoisomers. LCMS (m/z) 548.0, 550.0[M+H], Tr=2.68 min.

Compound 94d.1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-3-methyl-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide

To a solution of((S)-2-{3-[(R)-1-(7-bromo-quinolin-2-yl)-thylcarbamoyl]-3-methyl-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethyl)-carbamicacid tert-butyl ester (1.92 g, 3.5 mmol) in dichloromethane (15 mL) at0° C. was added trifluoroacetic acid (3 mL). The reaction mixture wasallowed to warm to room temperature and then stirred for 4 h. Thereaction mixture was concentrated and the residue was co-evaporated withtoluene (3×) to give1-((S)-2-amino-propionyl)-3-methyl-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide trifluoroacetate salt(3.5 mmol) as a brown oil. LCMS (m/z) 448.0, 450.0 [M+H], Tr=1.22 min.1-((S)-2-Amino-propionyl)-3-methyl-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide trifluoroacetate salt(3.5 mmol) was dissolved in dichloromethane (20 mL) and the solution wascooled to 0° C. (S)-2-Hydroxy-3-methyl-butyric acid (458 mg, 3.8 mmol),N,N-diisopropylethylamine (5 mL, 28.0 mmol) and(benzotriazol-1-yloxy)tris(dimethylamino) phosphoniumhexafluorophosphate (1.86 g, 4.2 mmol) were added. The reaction mixturewas allowed to warm to room temperature and was stirred for 16 h. Thesolvent was evaporated and the residue was dissolved in ethyl acetate.The organic solution was washed with citric acid (10%), saturated sodiumhydrogen carbonate solution and brine, dried over anhydrous magnesiumsulfate and the solvent was evaporated. The residue was purified bysilica gel chromatography eluting with a gradient of ethylacetate/methanol 1:0 to 20:1 to afford the title compound (745 mg, 39%over 2 steps) as a white foam and as a 1:4 mixture of diastereoisomers.LCMS (m/z) 548.0, 550.0 [M+H], Tr=2.21 min.

Compound 94e

A stirred solution of1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-3-methyl-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (164 mg, 0.3 mmol),4-oxo-1-vinyl-cyclohexanecarboxylic acid (50 mg, 0.3 mmol),tri(o-tolyl)phosphine (18 mg, 0.06 mmol) and triethylamine (91 mg, 0.13mL, 0.9 mmol) in 1,4-dioxane (10 mL) was degassed with nitrogen for 15min. The solution was heated to 50° C. andtris(dibenzylideneacetone)dipalladium(0) (28 mg, 0.03 mmol) was added.The reaction mixture was heated at 80° C. for 1 h. The reaction mixturewas cooled to room temperature and the mixture was filtered through ahydrophobic frit and the filtrate was evaporated to afford crude1-((E)-2-{2-[(R)-1-({1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-3-methyl-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-oxo-cyclohexanecarboxylicacid (0.3 mmol) as a yellow gum and as a mixture of diastereoisomers.LCMS (m/z) 636.2 [M+H], Tr=1.65 min. A solution of2-methyl-6-nitrobenzoic anhydride (206 mg, 0.6 mmol) and4-(dimethylamino)-pyridine (146 mg, 1.2 mmol) in dichloromethane (200mL) containing 4 Å molecular sieves (200 mg) was stirred at roomtemperature under nitrogen. A solution of crude1-((E)-2-{2-[(R)-1-({1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-3-methyl-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-4-oxo-cyclohexanecarboxylicacid (0.3 mmol) in dichloromethane (5 mL) was added dropwise over 4 hand the reaction mixture was stirred at room temperature for 18 h. Thereaction mixture was filtered through Celite and the solution waspartially evaporated to a volume of ˜30 mL. The solution was washed withice-cold saturated ammonium chloride solution, ice-cold saturated sodiumbicarbonate solution and brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography eluting with a gradient ofiso-hexanes/ethyl acetate 1:1 to 0:1 to afford the title compound (77mg, 41%) as a white solid and as a mixture of diastereoisomers. LCMS(m/z) 618.2 [M+H], Tr=2.38 min.

Compound 94

A solution of compound 94e (41 mg, 0.066 mmol) in anhydroustetrahydrofuran (4 mL) was stirred at −78° C. under nitrogen. Lithiumborohydride solution (2 M in tetrahydrofuran, 0.15 mL, 0.23 mmol) wasadded and the reaction mixture was stirred at −78° C. for 30 min.Hydrochloric acid (2 M) and ethyl acetate were added and the reactionmixture was warmed to room temperature. The organic layer was separatedand the aqueous layer was extracted with ethyl acetate. The organicextracts were combined and washed with brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theorange residue was purified by reverse phase preparative HPLC elutingwith acetonitrile (modified with 0.1% formic acid)/water (modified with0.1% formic acid) 3:7. Fractions containing the product were combinedand evaporated. The residue was dissolved in ethyl acetate and thesolution was washed with a small volume of saturated sodium hydrogencarbonate solution and brine. The organic solution was filtered througha hydrophobic frit and the filtrate was evaporated to afford the titlecompound (12 mg, 29%) as a white solid and as a 9:1 mixture ofdiastereoisomers. ¹H NMR (300 MHz, CD₃OD) (peaks for majordiastereoisomer) 61.04 (d, J=6.5 Hz, 3H), 1.10 (d, J=6.7 Hz, 3H),1.20-1.40 (m, 3H), 1.48 (s, 3H), 1.58 (d, J=7.1 Hz, 3H), 1.59 (d, J=7.3Hz, 3H), 1.65-1.80 (m, 3H), 1.95-2.25 (m, 4H), 2.34-2.38 (m, 1H),2.56-2.71 (m, 2H), 3.35-3.40 (m, 1H), 3.60-3.67 (m, 1H), 4.42-4.50 (m,1H), 5.14 (q, J=7.2 Hz, 1H), 5.27 (d, J=7.4 Hz, 1H), 5.74 (q, J=7.1 Hz,1H), 6.34 (d, J=16.3 Hz, 1H), 6.60 (d, J=16.3 Hz, 1H), 7.46 (d, J=8.5Hz, 1H), 7.58 (d, J=8.3 Hz, 1H), 7.84 (d, J=8.3 Hz, 1H), 7.93 (br s,1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 620.2 [M+H], Tr=2.17 min.

Example 95 Compound 95

Compound 95a. Hydroxy-oxetan-3-yl-acetic acid ethyl ester

To a stirred solution of oxetan-3-yl-acetic acid ethyl ester (obtainedfrom Activate Scientific GmbH), (1.04 g, 7.21 mmol) in anhydroustetrahydrofuran (72 mL) at −78° C. under nitrogen was added dropwisesodium hexamethyldisilazide (0.6 M in toluene, 14.4 mL, 8.66 mmol) togive a yellow orange solution. The reaction mixture was stirred at −78°C. for 30 min and then a solution of2-(phenylsulfonyl)-3-phenyl-oxaziridine (2.83 g, 10.8 mmol) intetrahydrofuran (7 mL) was added over 2 min. The reaction mixture wasstirred at −78° C. for 90 min and then saturated ammonium chloridesolution (10 mL) was added via syringe. The reaction mixture was allowedto warm to ambient temperature and after 5 min further saturatedammonium chloride solution (10 mL) was added and then stirred at ambienttemperature for 1 h. The reaction mixture was neutralised to pH 7 withhydrochloric acid (2 M) and the mixture diluted with water and extractedwith ethyl acetate (4×). The combined organic extracts were washed withbrine, dried over anhydrous magnesium sulfate, filtered and partiallyevaporated to about ˜10 mL. The mixture was diluted with dichloromethaneand the solids were filtered off, the filtrate was evaporated and theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/diethyl ether 1:0 to 1:3 to give the title compound (688 mg,60%) as a white gum. ¹H NMR (300 MHz, CDCl₃) δ1.30 (t, J=7.2 Hz, 3H),2.95 (d, J=5.1 Hz, 1H), 3.26-3.38 (m, 1H), 4.26 (q, J=7.1 Hz, 2H), 4.42(dd, J=7.4, 5.1 Hz, 1H), 4.67-4.81 (m, 4H). LCMS (m/z) 161.2 [M+H]Tr=0.72 min

Compound 95b. 2,2-Dimethyl-but-3-enoic acidethoxycarbonyl-oxetan-3-yl-methyl ester

To a stirred solution of hydroxy-oxetan-3-yl-acetic acid ethyl ester(330 mg, 2.06 mmol), 2-methyl-6-nitrobenzoic anhydride (1.42 g, 4.12mmol) and 4-dimethylamino pyridine (100 mg, 0.82 mmol) indichloromethane (4 mL) was added a solution of 2,2-dimethyl-but-3-enoicacid (294 mg, 2.58 mmol) in dichloromethane (4 mL). The reaction mixturewas stirred for 4 h and then evaporated. The residue was purified bysilica gel chromatography using iso-hexanes/diethyl ether 4:1 to givethe title compound (383 mg, 73%) as a white solid. ¹H NMR (300 MHz,CDCl₃) δ1.27 (t, J=7.1 Hz, 3H), 1.39 (s, 3H), 1.41 (s, 3H), 3.48-3.61(m, 1H), 4.20 (q, J=7.1 Hz, 2H), 4.62-4.68 (m, 2H), 4.72-4.85 (m, 2H),5.12-5.24 (m, 3H), 6.09 (dd, J=17.5, 10.6 Hz, 1H). LCMS (m/z)257.1[M+H], Tr=2.31 min

Compound 95c. 2,2-Dimethyl-but-3-enoic acid carboxy-oxetan-3-yl-methylester

To a stirred solution of 2,2-dimethyl-but-3-enoic acidethoxycarbonyl-oxetan-3-yl-methyl ester (207 mg, 0.808 mmol) intetrahydrofuran (81 mL) was added a solution of potassium carbonate (123mg, 0.888 mmol) in water (20.25 mL) followed by methanol (4.05 mL). Thereaction mixture was heated to reflux for 7 h and then allowed to cool.Hydrochloric acid (2 M, 400 μL) was added (pH 7) and the solvents wereevaporated. The residue was extracted with dichloromethane (2×) and thewashes discarded. The aqueous was acidified with hydrochloric acid (2 M)to pH 5, extracted with dichloromethane (2×) and the combined organicextracts passed through a hydrophobic frit and evaporated. To theaqueous layer was added hydrochloric acid (about 5 drops) and themixture was extracted with dichloromethane (2×) and the combined organicextracts passed through a hydrophobic frit, evaporated and combined withprevious material to give the title compound (123 mg, 67%) as ayellow/green oil. ¹H NMR (300 MHz, CDCl₃) δ1.39 (s, 3H), 1.40 (s, 3H),3.54-3.67 (m, 1H), 4.67-4.75 (m, 2H), 4.81-4.92 (m, 2H), 5.14 (d, J=10.9Hz, 1H), 5.15-5.32 (m, 2H), 6.08 (dd, J=17.5, 10.5 Hz, 1H), 9.30 (br s,1H). LCMS (m/z) 229.1 [M+H] Tr=1.67 min.

Compound 95d.(S)-1-((S)-2-Amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid

To a stirred solution of(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (330 mg, 0.618 mmol) indichloromethane (10 mL) at 0° C. was added trifluoroacetic acid (3.3 mL)and the reaction mixture stirred for 1 hour and then evaporated. Theresidue was dissolved in dichloromethane and washed with saturatedbicarbonate solution (2×), the aqueous extracts washed withdichloromethane and the combined organic extracts passed through ahydrophobic frit and evaporated to give a yellow foam (232 mg) that wascarried through to the next step directly.

Compound 95e. 2,2-Dimethyl-but-3-enoic acid((S)-2-{(S)-3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethylcarbamoyl)-oxetan-3-yl-methylester

A stirred solution of(S)-1-((S)-2-amino-propionyl)-hexahydro-pyridazine-3-carboxylic acid[(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (141 mg, 0.618 mmol) and2,2-dimethyl-but-3-enoic acid carboxy-oxetan-3-yl-methyl ester (315 mg,0.725 mmol) in anhydrous N,N-dimethylformamide (10 mL) was cooled to 0°C. under nitrogen. 2,2,6,6-Tetramethylpiperidine (261 mg, 313 μL, 1.85mmol) was added, rapidly followed by(1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate (371 mg, 0.865 mmol). The reaction mixture wasallowed to warm to room temperature and stirred for 15 h. The reactionmixture was diluted with dichloromethane and washed with saturatedammonium chloride solution (2×), saturated sodium bicarbonate solution(2×), the organic layer was filtered through a phase-separator andevaporated. The residue was purified by silica gel chromatography usinga gradient of iso-hexanes/ethyl acetate 1:0 to 0:1 to give the titlecompound (167 mg, 42%) as a yellow/brown foam and as an approximate 1:1mixture of diastereoisomers. LCMS (m/z) 644.2, 646.1[M+H], Tr=2.63 min.

Compound 95

A solution of 2,2-dimethyl-but-3-enoic acid((S)-2-{(S)-3-[(R)-1-(7-bromo-quinolin-2-yl)-ethylcarbamoyl]-tetrahydro-pyridazin-1-yl}-1-methyl-2-oxo-ethylcarbamoyl)-oxetan-3-yl-methylester (167 mg, 0.259 mmol), triethylamine (79 mg, 108 μL, 0.777 mmol)and tri(o-tolyl)phosphine (16 mg, 0.052 mmol) in 1,4-dioxane (86 mL) wasdegassed with nitrogen for 10 min and then warmed to 50° C. undernitrogen with stirring. Tris(dibenzylideneacetone)dipalladium(0) (24 mg,0.026 mmol) was added and the reaction mixture was heated to 80° C. for90 min and then more tri(o-tolyl)phosphine (32 mg, 0.104 mmol) andtris(dibenzylideneacetone)dipalladium(0) (48 mg, 0.052 mmol) were added.The reaction mixture was heated to 80° C. for 2 h, and moretri(o-tolyl)phosphine (16 mg, 0.52 mmol) andtris(dibenzylideneacetone)dipalladium(0) (24 mg, 0.026 mmol) were added.After heating at 80° C. for 1.5 h, more tri(o-tolyl)phosphine (16 mg,0.52 mmol) and tris(dibenzylideneacetone)dipalladium(0) (24 mg, 0.026mmol) were added. After heating at 80° C. for 30 min, the mixture wascooled and filtered over Celite and the filtrate was evaporated. Theresidue was purified by reverse phase preparative HPLC using a gradientof acetonitrile/water 1:9 to 7:3 to give the title compound (15.6 mg,11%) as a yellow solid. ¹H NMR (300 MHz, CD₃OD) δ1.46 (s, 3H), 1.50-1.73(m, 1H), 1.56 (s, 3H), 1.59 (d, J=6.7 Hz, 3H), 1.64 (d, J=7.2 Hz, 3H),1.90-2.01 (m, 1H), 2.23-2.32 (m, 1H), 2.63-2.74 (m, 1H), 3.47-3.62 (m,2H), 4.37-4.45 (m, 1H), 4.64 (t, J=6.5 Hz, 1H), 4.72-4.82 (m, 4H), 5.09(q, J=6.7 Hz, 1H), 5.75 (q, J=7.4 Hz, 1H), 5.82 (d, J=6.9 Hz, 1H), 6.38(d, J=16.3 Hz, 1H), 6.60 (d, J=16.3 Hz, 1H), 7.45 (d, J=8.5 Hz, 1H),7.61 (dd, J=8.5, 1.6 Hz, 1H), 7.81 (br s, 1H), 7.85 (d, J=8.5 Hz, 1H),8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 564.2 [M+H], Tr=2.16 min.

Example 96 Compound 96

A cooled (0° C.) solution of compound 45 (63.0 mg, 0.091 mmol) indichloromethane (10 mL) was treated with a solution of hydrogen chloride(4 M) in 1,4-dioxane (0.12 mL, 0.455 mmol). After stirring at roomtemperature for 6 days, dichloromethane (10 mL) and hydrogen chloride (4M) in 1,4-dioxane (0.12 mL, 0.455 mmol) were added. After stirring for 4h at room temperature, the volatiles were removed in vacuo and theresidual water was azeotroped off with toluene to provide a white solidwhich was suspended in dichloromethane (5 mL) and the mixture wassubsequently treated with triethylamine (0.02 mL, 0.136 mmol) andacetaldehyde (0.01 mL, 0.182 mmol). After stirring at room temperaturefor 30 min, the reaction mixture was treated with sodiumtriacetoxyborohydride (38.6 mg, 0.182 mmol). After stirring for 1 h atroom temperature the reaction mixture was quenched with a few drops ofsaturated sodium bicarbonate and filtered through a phase separator. Thevolatiles were removed in vacuo and the residue was purified by reversephase preparative HPLC using a gradient of acetonitrile/water 5:95 to3:7 modified with 0.1% formic acid. After evaporation of the volatiles,the white solid was partitioned between ethyl acetate and a saturatedsolution of sodium bicarbonate. The organics were filtered through aphase separator and the volatiles were removed in vacuo to provide thetitle compound (8.3 mg, 15% over 2 steps) as a white solid. ¹H NMR (300MHz, CD₃OD): δ1.02-1.18 (m, 9H), 1.22-1.33 (m, 1H), 1.51-1.73 (m, 7H),1.89-2.00 (m, 1H), 2.11 (d, J=11.6 Hz, 1H), 2.16-2.36 (m, 3H), 2.39-2.57(m, 2H), 2.61-2.73 (m, 1H), 2.74-2.82 (m, 1H), 3.54-3.67 (m, 2H),3.87-4.02 (m, 2H), 4.38-4.47 (m, 1H), 4.99 (d, J=12.0 Hz, 1H), 5.05-5.14(m, 2H), 5.79 (q, J=7.4 Hz, 1H), 6.13 (d, J=16.3 Hz, 1H), 6.92 (d,J=16.3 Hz, 1H), 7.46 (d, J=8.5 Hz, 1H), 7.60 (dd, J=8.5, 1.6 Hz, 1H),7.86 (d, J=8.5 Hz, 1H), 7.91 (s, 1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z)621.3 [M+H], Tr=1.20 min.

Example 97 Compound 97

A cooled (0° C.) solution of compound 45 (85.3 mg, 0.123 mmol) indichloromethane (10 mL) was treated with a solution of hydrogen chloride(4 M) in 1,4-dioxane (0.31 mL, 1.231 mmol). After stirring at roomtemperature for 2 days, more hydrogen chloride (4 M) in 1,4-dioxane (0.3mL, 1.2 mmol) was added. After stirring for 1.5 h at room temperature,the volatiles were removed in vacuo and the residual water wasazeotroped off with toluene to provide a white solid which was suspendedin dichloromethane (5 mL) and the mixture was subsequently treated withtriethylamine (0.04 mL, 0.246 mmol) and methanesulfonyl chloride (0.01mL, 0.148 mmol). After stirring at room temperature for 30 min, thereaction mixture was treated with sodium triacetoxyborohydride (38.6 mg,0.182 mmol). After stirring for 1 h at room temperature the reactionmixture was quenched with a saturated sodium bicarbonate solution. Theaqueous layer was extracted with dichloromethane. The organics werecombined and filtered through a phase separator. The volatiles wereremoved in vacuo and the residue was purified by reverse phasepreparative HPLC using a gradient of acetonitrile/water 5:95 to 1:0 toprovide the title compound (14.7 mg, 18% over 2 steps) as a white solid.¹H NMR (300 MHz, CD₃OD) δ1.05-1.12 (m, 6H), 1.52-1.73 (m, 8H), 1.91-2.00(m, 1H), 2.17-2.30 (m, 2H), 2.62-2.74 (m, 1H), 2.93 (s, 3H), 3.01 (d,J=11.8 Hz, 1H), 3.05-3.13 (m, 1H), 3.51-3.68 (m, 2H), 3.83-3.94 (m, 1H),4.07-4.15 (m, 1H), 4.24-4.31 (m, 1H), 4.39-4.47 (m, 1H), 5.02 (d, J=11.8Hz, 1H), 5.10 (q, J=6.7 Hz, 1H), 5.19 (d, J=8.7 Hz, 1H), 5.81 (q, J=7.3Hz, 1H), 6.19 (d, J=16.3 Hz, 1H), 6.99 (d, J=16.3 Hz, 1H), 7.48 (d,J=8.5 Hz, 1H), 7.62 (dd, J=8.5, 1.6 Hz, 1H), 7.87 (d, J=8.5 Hz, 1H),7.93 (s, 1H), 8.27 (d, J=8.2 Hz, 1H). LCMS (m/z) 671.3 [M+H], Tr=2.07min.

Example 98 Compound 98

Compound 98a. 2,2-Bis-hydroxymethyl-3-oxo-butyric acid tert-butyl ester

A solution of 3-oxo-butyric acid tert-butyl ester (15.8 g, 100 mmol) andaqueous formaldehyde (37% solution in water, 18 mL, 230 mmol) in1,4-dioxane (50 mL) was stirred at 0° C. under nitrogen. A solution oftriethylamine (505 mg, 0.7 mL, 5 mmol) in tetrahydrofuran (5 mL) wasadded dropwise and the reaction mixture was stirred at 40° C. for 1 hand then at room temperature for 20 h. The majority of the solvent wasevaporated, water was added and the mixture was extracted with ethylacetate. The organic extracts were combined and washed with brine. Theorganic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 7:3 to 1:1to afford 2,2-bis-hydroxymethyl-3-oxo-butyric acid tert-butyl ester(8.67 g, 40%) as a clear oil. ¹H NMR (300 MHz, d₆-DMSO) 61.38 (s, 9H),2.07 (s, 3H), 3.81-3.86 (m, 4H), 4.77 (t, J=5.1 Hz, 2H). LCMS (m/z)217.0 [M−H], Tr=1.45 min.

Compound 98b. 5-Acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acidtert-butyl ester

A mixture of 2,2-bis-hydroxymethyl-3-oxo-butyric acid tert-butyl ester(8.67 g, 40 mmol), acetone (30 mL, 400 mmol), 2,2-dimethoxypropane (41.6g, 50 mL, 400 mmol) and 4-toluenesulfonic acid hydrate (152 mg, 0.8mmol) was stirred at room temperature for 18 h. The solvent waspartially evaporated to a volume of ˜20 mL. Saturated sodium hydrogencarbonate solution was added and the mixture was extracted with ethylacetate. The organic extracts were combined, washed with water andbrine. The organic layer was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 9:1 to 4:1to afford the title compound (7.82 g, 75%) as a clear oil. ¹H NMR (300MHz, CDCl₃) δ1.39 (s, 3H), 1.44 (s, 3H), 1.48 (s, 9H), 2.32 (s, 3H),6.36 (ABq, Δδ_(AB)=0.05, J_(AB)=11.8 Hz, 4H). LCMS (m/z) 281.2 [M+Na],Tr=2.44 min.

Compound 98c. 2,2-Dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acidtert-butyl ester

A solution of N,N-diisopropylamine (1.51 g, 2.1 mL, 15 mmol) inanhydrous tetrahydrofuran (30 mL) was stirred at −78° C. under nitrogen.n-Butyl lithium (2.5 M in hexanes, 5.6 mL, 14 mmol) was added dropwiseand the reaction mixture was stirred at −78° C. for 30 min. A solutionof 5-acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acid tert-butyl ester(2.58 g, 10 mmol) in tetrahydrofuran (5 mL) was added and the reactionmixture was stirred at −78° C. for 15 min. A solution ofN-phenyl-(bis-trifluoromethanesulfonamide) (3.93 g, 11 mmol) intetrahydrofuran (30 mL) was added and the reaction mixture was stirredat −78° C. for 15 min. The cooling bath was removed and the reactionmixture was stirred at room temperature for 30 min. The solvent wasevaporated and diethyl ether (50 mL) was added. The solution was cooledto 5° C. and was washed with cold sodium hydroxide solution (1 M, 3×30mL) and brine. The organic solution was separated, dried over anhydrousmagnesium sulfate, filtered and the solvent evaporated to afford2,2-dimethyl-5-(1-trifluoromethanesulfonyloxy-vinyl)-[1,3]dioxane-5-carboxylicacid tert-butyl ester (3.84 g, 10 mmol) as a yellow oil. A solution of2,2-dimethyl-5-(1-trifluoromethanesulfonyloxy-vinyl)-[1,3]dioxane-5-carboxylicacid tert-butyl ester (3.84 g, 10 mmol) and tri-n-butylamine (5.55 g, 7mL, 30 mmol) in N,N-dimethylformamide (20 mL) was stirred at roomtemperature under nitrogen. Bis(triphenylphosphine)palladium(II)dichloride (350 mg, 0.5 mmol) and formic acid (920 mg, 0.77 mL, 20 mmol)were added and the reaction mixture was heated at 60° C. for 90 min. Thereaction mixture was cooled to room temperature and ethyl acetate andwater were added. The organic extracts were washed with water (5×) andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using iso-hexanes/ethyl acetate 9:1 followed by silicagel chromatography using a gradient of iso-hexanes/ethyl acetate 19:1 to9:1 to afford the title compound (1.09 g, 45%) as a clear oil. ¹H NMR(300 MHz, CDCl₃) δ 1.41 (s, 3H), 1.45 (s, 3H), 1.50 (s, 9H), 3.82 (d,J=11.8 Hz, 2H), 4.25 (d, J=11.8 Hz, 2H), 5.25 (d, J=17.2 Hz, 1H), 5.26(d, J=10.7 Hz, 1H), 5.73 (dd, J=17.2, 11.7 Hz, 1H).

Compound 98d. 2,2-Bis-hydroxymethyl-but-3-enoic acid tert-butyl ester

A solution of 2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acidtert-butyl ester (969 mg, 4 mmol) and 4-toluenesulfonic acid hydrate (76mg, 0.4 mmol) in methanol (10 mL) was stirred at room temperature for 30min. The solvent was evaporated and the residue was purified by silicagel chromatography using a gradient of iso-hexanes/ethyl acetate 4:1 to1:1 to afford the title compound (660 mg, 82%) as a white solid. ¹H NMR(300 MHz, d₆-DMSO): δ1.38 (s, 9H), 3.54-3.68 (m, 4H), 4.56 (t, J=5.4 Hz,2H), 5.12 (d, J=10.5 Hz, 1H), 5.14 (d, J=18.3 Hz, 1H), 5.80 (dd, J=18.3,10.5 Hz, 1H). LCMS (m/z) 225.2 [M+Na], Tr=1.47 min.

Compound 98e. 2-Oxo-5-vinyl-[1,3]dioxane-5-carboxylic acid tert-butylester

A solution of 2,2-bis-hydroxymethyl-but-3-enoic acid tert-butyl ester(202 mg, 1 mmol) and pyridine (0.5 mL, 6 mmol) in dichloromethane (5 mL)was stirred at −78° C. under nitrogen. A solution of triphosgene (150mg, 0.5 mmol) in dichloromethane (2.5 mL) was added dropwise and thereaction mixture was stirred at −78° C. for 1 h. The reaction mixturewas warmed to room temperature and stirred at room temperature for 15min and saturated ammonium chloride solution was added. The organiclayer was separated, washed with hydrochloric acid (1 M), saturatedsodium hydrogen carbonate solution and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 4:1 to 3:2 to afford the title compound (204mg, 89%) as a clear gum. ¹H NMR (300 MHz, CDCl₃): δ 1.48 (s, 9H), 4.38(d, J=10.7 Hz, 2H), 4.71 (d, J=10.7 Hz, 2H), 5.39 (d, J=17.4 Hz, 1H),5.47 (d, J=10.7 Hz, 1H), 5.77 (dd, J=17.4, 10.7 Hz, 1H). LCMS (m/z)229.1 [M+H], Tr=2.04 min.

Compound 98f. 2-Oxo-5-vinyl-[1,3]dioxane-5-carboxylic acid

A solution of 2-oxo-5-vinyl-[1,3]dioxane-5-carboxylic acid tert-butylester (200 mg, 0.88 mmol) in dichloromethane (4 mL) was stirred at 0° C.Trifluoroacetic acid (2 mL) was added and the reaction mixture wasstirred at 0° C. for 30 min and then at room temperature for 3 h. Thesolvent was evaporated and the residue was triturated with diethylether/iso-hexanes (1:1) to afford the title compound (135 mg, 90%) as awhite solid. ¹H NMR (300 MHz, d₆-DMSO) 64.51 (d, J=10.5 Hz, 2H), 4.67(d, J=10.5 Hz, 2H), 5.38 (d, J=17.6 Hz, 1H), 5.43 (d, J=10.7 Hz, 1H),5.83 (dd, J=17.6, 10.7 Hz, 1H), 13.5-14.0 (br s, 1H). LCMS (m/z) 173.2[M+H], Tr=0.62 min.

Compound 98

A stirred solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (164 mg, 0.3 mmol),2-oxo-5-vinyl-[1,3]dioxane-5-carboxylic acid (57 mg, 0.33 mmol),tri(o-tolyl)phosphine (18 mg, 0.06 mmol) and triethylamine (91 mg, 0.13mL, 0.9 mmol) in 1,4-dioxane (10 mL) was degassed with nitrogen for 15min. The solution was heated to 50° C. andtris(dibenzylideneacetone)dipalladium(0) (28 mg, 0.03 mmol) was added.The reaction mixture was heated at 80° C. for 45 min. The reactionmixture was cooled to room temperature and the mixture was filteredthrough a hydrophobic frit and the filtrate was evaporated to affordcrude5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-2-oxo-[1,3]dioxane-5-carboxylicacid (0.3 mmol) as a yellow gum. LCMS (m/z) 626.2 [M+H], Tr=1.67 min. Asolution of 2-methyl-6-nitrobenzoic anhydride (206 mg, 0.6 mmol) and4-(dimethylamino)-pyridine (146 mg, 1.2 mmol) in dichloromethane (200mL) containing 4 Å molecular sieves (200 mg) was stirred at roomtemperature under nitrogen. A solution of crude5-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-2-oxo-[1,3]dioxane-5-carboxylicacid (0.3 mmol) in dichloromethane (5 mL) was added dropwise over 4 hand the reaction mixture was stirred at room temperature for 16 h. Thereaction mixture was filtered through Celite and the solution waspartially evaporated to a volume of ˜50 mL. The solution was washed withice-cold saturated ammonium chloride solution, ice-cold saturated sodiumbicarbonate solution and brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography eluting with a gradient ofethyl acetate/methanol 1:0 to 5:1 then by reverse phase preparative HPLCeluting with acetonitrile/water 7:13. Fractions containing the productwere combined and the majority of the organic solvent was evaporated.The resulting aqueous solution was extracted with dichloromethane. Theorganic extracts were combined, filtered through a hydrophobic frit andthe filtrate was evaporated to afford the title compound (3.1 mg, 2%) asa white solid. ¹H NMR (300 MHz, CD₃CN): δ0.96 (d, J=6.7 Hz, 3H), 1.04(d, J 15=6.7 Hz, 3H), 1.41-1.49 (m, 2H), 1.54 (d, J=6.7 Hz, 3H), 1.55(d, J=7.1 Hz, 3H), 1.59-1.64 (m, 1H), 1.88-1.93 (m, 1H), 2.19-2.24 (m,1H), 2.56-2.64 (m, 1H), 3.48-3.57 (m, 1H), 4.19 (d, J=12.0 Hz, 1H),4.38-4.43 (m, 1H), 4.59 (d, J=10.9 Hz, 1H), 4.71 (d, J=10.9 Hz, 1H),4.85-4.91 (m, 2H), 5.06-5.10 (m, 1H), 5.23 (d, J=7.8 Hz, 1H), 5.73-5.79(m, 1H), 6.26 (d, J=16.5 Hz, 1H), 6.83 (d, J=16.5 Hz, 1H), 7.18 (d,J=8.0 Hz, 1H), 7.48 (d, J=8.5 Hz, 1H), 7.64 (dd, J=8.5, 1.6 Hz, 1H),7.90 (d, J=8.5 Hz, 1H), 7.99 (br s, 1H), 8.28 (d, J=8.5 Hz, 1H), 8.85(br d, J=5.0 Hz, 1H). LCMS (m/z) 608.2 [M+H], Tr=2.01 min.

Example 99 Compound 99

Compound 99a. 5-Acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acid ethylester

Compound 99a was prepared in the same manner as5-acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acid tert-butyl esterusing ethyl acetoacetate instead of 3-oxo-butyric acid tert-butyl esterin 31% yield over 2 steps. ¹H NMR (300 MHz, CDCl₃) δ1.29 (t, J=7.1 Hz,3H), 1.40 (s, 3H), 1.44 (s, 3H), 2.33 (s, 3H), 4.23 (q, J=7.1 Hz, 2H),4.30 (ABq, Δδ_(AB)=0.04, J_(AB)=11.9 Hz, 4H).

Compound 99b. 2,2-Dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid ethylester

Compound 99b was prepared in the same manner as2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid tert-butyl esterusing 5-acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acid ethyl esterinstead of 5-acetyl-2,2-dimethyl-[1,3]dioxane-5-carboxylic acidtert-butyl ester in 50% yield over 2 steps. ¹H NMR (300 MHz, CDCl₃)δ1.31 (t, J=7.1 Hz, 3H), 1.41 (s, 3H), 1.46 (s, 3H), 4.10 (ABq,Δδ_(AB)=0.44, J_(AB)=11.9 Hz, 4H), 4.26 (q, J=7.1 Hz, 2H), 5.24 (d,J=17.4 Hz, 1H), 5.28 (d, J=10.9 Hz, 1H), 5.24 (dd, J=17.4, 10.9 Hz, 1H).LCMS (m/z) 237.0 [M+Na], Tr=2.14 min.

Compound 99c. 2,2-Bis-hydroxymethyl-but-3-enoic acid ethyl ester

Compound 99c was prepared in the same manner as2,2-bis-hydroxymethyl-but-3-enoic acid tert-butyl ester using2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid ethyl ester insteadof 2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid tert-butyl esterin 78% yield. ¹H NMR (300 MHz, d₆-DMSO) 61.17 (t, J=7.0 Hz, 3H),3.59-3.71 (m, 4H), 4.06 (q, J=7.0 Hz, 2H), 4.65 (t, J=5.6 Hz, 2H), 5.13(d, J=18.5 Hz, 1H), 5.15 (d, J=10.5 Hz, 1H), 5.80 (dd, J=18.5, 10.5 Hz,1H). LCMS (m/z) 197.1 [M+Na], Tr=0.96 min.

Compound 99d. 2,2-Bis-fluoromethyl-but-3-enoic acid

To a solution of 2,2-bis-hydroxymethyl-but-3-enoic acid ethyl ester (338mg, 1.9 mmol) and 2,6-lutidine (813 mg, 0.88 mL, 7.6 mmol) indichloromethane (10 mL) at −78° C. under nitrogen was added dropwisetrifluoromethanesulfonic anhydride (1.07 g, 0.66 mL, 3.8 mmol) and thereaction mixture was warmed to room temperature and stirred at roomtemperature for 30 min. The solution was washed with ice-coldhydrochloric acid (1 M) and brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated to affordcrude 2,2-bis-trifluoromethanesulfonyloxymethyl-but-3-enoic acid ethylester (1.9 mmol) as an orange oil which was used immediately in the nextstep. A solution of2,2-bis-trifluoromethanesulfonyloxymethyl-but-3-enoic acid ethyl ester(1.9 mmol) in tetrahydrofuran (5 mL) was stirred at 0° C. undernitrogen. Tetra-n-butylammonium fluoride (1 M in tetrahydrofuran, 4.75mL, 4.75 mmol) was added and the reaction mixture was stirred at 0° C.for 30 min and then at room temperature for 2 h. The solvent wasevaporated. The residue was dissolved in dichloromethane and thesolution was washed with saturated sodium hydrogen carbonate solution,water and brine. The organic solution was filtered through a hydrophobicfrit and the filtrate was evaporated. The residue was purified by silicagel chromatography using a gradient of pentane/diethyl ether 19:1 to 4:1to afford 2,2-bis-fluoromethyl-but-3-enoic acid ethyl ester (1.9 mmol)contaminated with solvent as a clear oil. A solution of crude2,2-bis-fluoromethyl-but-3-enoic acid ethyl ester (1.9 mmol) intetrahydrofuran (10 mL) was stirred at 0° C. under nitrogen. A solutionof lithium hydroxide monohydrate (240 mg, 5.7 mmol) in water (2.5 mL)was added and the reaction mixture was stirred at 0° C. for 1 h and thenat room temperature for 24 h. The reaction mixture was acidified to pH 2with hydrochloric acid (2 M). Water was added and the mixture wasextracted with diethyl ether. The organic extracts were combined, washedwith brine. The organic solution was filtered through a hydrophobic fritand the filtrate was evaporated to afford the title compound (400 mg) asa clear oil. ¹H NMR (300 MHz, d₆-DMSO) δ 4.54-4.83 (m, 4H), 5.33 (d,J=17.8 Hz, 1H), 5.36 (d, J=10.7 Hz, 1H), 5.81 (dd, J=17.8, 10.7 Hz, 1H),13.0-13.5 (br s, 1H). LCMS (m/z) 149.2 [M−H], Tr=1.45 min.

Compound 99e.(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-tert-Butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester

A solution of [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-carbamic acidtert-butyl ester (105 mg, 0.3 mmol), 2,2-bis-fluoromethyl-but-3-enoicacid (50 mg, 0.33 mmol), tri(o-tolyl)phosphine (18 mg, 0.06 mmol) andtriethylamine (91 mg, 0.13 mL, 0.9 mmol) in 1,4-dioxane (3 mL) wasdegassed with nitrogen for 15 min.Tris(dibenzylideneacetone)dipalladium(0) (28 mg, 0.03 mmol) was addedand the reaction mixture was heated to 60° C. for 1 h. The reactionmixture was cooled to room temperature and the mixture was filteredthrough a hydrophobic frit and the filtrate was evaporated to affordcrude(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoicacid (0.3 mmol) as a yellow gum. LCMS (m/z) 421.1 [M+H], Tr=2.27 min. Asolution of crude(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoicacid (0.3 mmol), N,N-diisopropylethylamine (96 mg, 0.13 mL, 0.75 mmol),4-(dimethylamino)-pyridine (73 mg, 0.6 mmol) and 2-methyl-6-nitrobenzoicanhydride (193 mg, 0.56 mmol) in anhydrous dichloromethane (5 mL) wasstirred at 0° C. under nitrogen. A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (142 mg, 0.33 mmol) in dichloromethane(2 mL) was added and the reaction mixture was stirred at 0° C. for 30min and then at room temperature for 18 h. The reaction mixture wasdiluted with dichloromethane and the solution was washed with ice-coldsaturated sodium hydrogen carbonate solution, water, ice-coldhydrochloric acid (1 M), water and brine. The organic solution wasfiltered through a hydrophobic frit and the filtrate was evaporated. Theresidue was purified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 2:1 to 0:1 to afford the partially purifiedtitle compound (26 mg, 10%) as a yellow gum. LCMS (m/z) 834.3, 836.2[M+H], Tr=3.39 min.

Compound 99

A solution of(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid 2,2,2-trichloro-ethyl ester (100 mg, 0.11 mmol) in tetrahydrofuran(3 mL) was stirred at 0° C. under nitrogen. An ice-cold aqueous solutionof sodium hydroxide (0.1 M, 1.1 mL, 0.11 mmol) was added and thereaction mixture was stirred at 0° C. for 20 min. Cold hydrochloric acid(1 M) was added to acidify the mixture to pH 2 and the solvent wasevaporated. The residue was co-evaporated with tetrahydrofuran/toluene(1:1, 3×) and the residue was triturated with diethyl ether (2×) and theresulting solid was dried to afford(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.1 mmol) as a yellow solid which was used crude in the nextreaction. LCMS (m/z) 704.3 [M+H], Tr=2.61 min. A mixture of crude(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-tert-butoxycarbonylamino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid (0.1 mmol) in hydrochloric acid (4 M in 1,4-dioxane, 2 mL) wasstirred at room temperature for 30 min. The solvent was evaporated andthe residue was co-evaporated with diethyl ether (2×) and the resultingsolid was dried to afford(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.1 mmol) as an off-white solid which was used crudein the next reaction. LCMS (m/z) 604.3 [M+H], Tr=1.53 min. A suspensionof crude(S)-1-[(S)-2-((S)-2-{(E)-4-[2-((R)-1-amino-ethyl)-quinolin-7-yl]-2,2-bis-fluoromethyl-but-3-enoyloxy}-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid hydrochloride (0.1 mmol) in dichloromethane (100 mL) was stirred at0° C. under nitrogen. A solution of N,N-diisopropylethylamine (52 mg,0.07 mL, 0.4 mmol) in dichloromethane (5 mL) was added and the resultingsolution was stirred at 0° C.2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (76 mg, 0.2 mmol) was added and thereaction mixture was stirred at 0° C. for 30 min and then at roomtemperature for 18 h. The solvent was partially evaporated to a volumeof ˜30 mL. The solution was washed with ice-cold saturated sodiumhydrogen carbonate solution, ice-cold hydrochloric acid (1 M) and brine.The organic solution was filtered through a hydrophobic frit and thefiltrate was evaporated. The residue was purified by silica gelchromatography eluting with a gradient of iso-hexanes/ethyl acetate 1:1to 0:1 followed silica gel chromatography eluting withiso-hexanes/acetone 3:2. The residue was triturated with diethyl etherand the resulting solid was dried to afford the title compound (4 mg, 7%over 3 steps) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.01 (d, J=6.5Hz, 3H), 1.09 (d, J=6.9 Hz, 3H), 1.49-1.55 (m, 2H), 1.61 (d, J=6.9 Hz,3H), 1.64 (d, J=7.1 Hz, 3H), 1.93-1.98 (m, 1H), 2.15-2.28 (m, 2H),2.68-2.76 (m, 1H), 3.59-3.63 (m, 1H), 4.41-4.46 (m, 1H), 4.70-5.13 (m,5H), 5.35 (d, J=8.9 Hz, 1H), 5.84 (q, J=7.1 Hz, 1H), 6.27 (d, J=16.5 Hz,1H), 6.81 (d, J=16.5 Hz, 1H), 7.49 (d, J=7.9 Hz, 1H), 7.70 (dd, J=8.5,1.6 Hz, 1H), 7.82 (br s, 1H), 7.88 (d, J=8.5 Hz, 1H), 8.28 (d, J=8.5 Hz,1H). LCMS (m/z) 586.2 [M+H], Tr=2.48 min.

Examples 100 and 101. Compounds 100 and 101

Compound 100a. 2-Allyloxy-2-methyl-propionic acid ethyl ester

A solution of 2-hydroxy-2-methyl-propionic acid ethyl ester (2.64 g, 2.7mL, 20 mmol) in N,N-dimethylformamide (20 mL) was stirred at 0° C. undernitrogen. Sodium hydride (60% dispersion in oil, 880 mg, 22 mmol) wasadded and the reaction mixture was stirred at 0° C. for 5 min. Allylbromide (2.18 g, 1.6 mL, 18 mmol) was added and the reaction mixture wasstirred at 0° C. for 30 min and then at room temperature for 18 h. Thereaction mixture was cooled to 0° C. and saturated ammonium chloridesolution was cautiously added to quench the reaction. Diethyl ether wasadded and the organic layer was separated, washed with water (3×) andbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 12:1 to 4:1 toafford the title compound (2.33 g, 67%) as an clear oil. ¹H NMR (300MHz, CDCl₃) δ 1.31 (t, J=7.1 Hz, 3H), 1.47 (s, 6H), 3.95-3.98 (m, 2H),4.21 (q, J=7.1 Hz, 2H), 5.15-5.20 (m, 1H), 5.27-5.39 (m, 1H), 5.90-6.03(m, 1H). LCMS (m/z) 195.2 [M+Na], Tr=2.24 min.

Compound 100b. (5,5-Dimethyl-[1,4]dioxan-2-yl)-methanol

A solution of 2-allyloxy-2-methyl-propionic acid ethyl ester (1.72 g, 10mmol) in anhydrous tetrahydrofuran (40 mL) was stirred at roomtemperature under nitrogen. Lithium borohydride (44 mg, 20 mmol) wasadded portion-wise and the reaction mixture was stirred at roomtemperature for 20 h. The reaction mixture was cooled to 0° C. andsaturated ammonium chloride solution was cautiously added followed byhydrochloric acid (2 M) to acidify the reaction mixture to pH 2. Themixture was extracted with diethyl ether and the organic extracts werecombined and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated to afford2-allyloxy-2-methyl-propan-1-ol (10 mmol) which was used crude in thenext step. LCMS (m/z) 153.2 [M+Na], Tr=1.16 min. A suspension of2-allyloxy-2-methyl-propan-1-ol (10 mmol) in dichloromethane (40 mL) wasstirred at 0° C. Meta-chloroperbenzoic acid (70% pure containing 20%water and 10% meta-chlorobenzoic acid, 2.9 g, 12 mmol) was added and thereaction mixture was stirred at 0° C. for 30 min and then at roomtemperature for 24 h. The reaction mixture was cooled to 0° C. andsaturated sodium hydrogen carbonate solution and sodium thiosulfatesolution were added and the mixture was stirred at 0° C. for 15 min. Theorganic layer was separated and the aqueous layer was extracted withdichloromethane (4×). The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated to afford2-methyl-2-oxiranylmethoxy-propan-1-ol (10 mmol) which was used crude inthe next step. LCMS (m/z) 169.2 [M+Na], Tr=0.72 min. A solution of2-methyl-2-oxiranylmethoxy-propan-1-ol (10 mmol) in dichloromethane (50mL) and racemic camphor-10-sulfonic acid (765 mg, 3.3 mmol) was stirredat room temperature for 18 h. Saturated sodium hydrogen carbonatesolution was added and the organic layer separated. The aqueous layerwas extracted with dichloromethane and the organic extracts werecombined, filtered through a hydrophobic frit and the filtrate wasevaporated. The residue was purified by silica gel chromatography usinga gradient of pentane/diethyl ether 2:3 to 1:4 to afford the titlecompound (826 mg, 57% over 3 steps) as a yellow oil. ¹H NMR (300 MHz,d₆-DMSO): δ1.02 (s, 3H), 1.20 (s, 3H), 3.23 (d, J=11.1 Hz, 1H),3.30-3.50 (m, 6H), 4.68 (t, J=5.4 Hz, 1H). LCMS (m/z) 169.2 [M+Na],Tr=0.72 min.

Compound 100c. 5,5-Dimethyl-[1,4]dioxane-2-carboxylic acid benzyl ester

A solution of (5,5-dimethyl-[1,4]dioxan-2-yl)-methanol (657 mg, 4.5mmol) in 1 M potassium hydroxide solution (6.7 mL, 6.7 mmol) was stirredat 5° C. A solution of potassium permanganate (1.42 g, 9 mmol) in water(10 mL) was added dropwise maintaining the internal temperature below10° C. The reaction mixture was stirred at 5° C. for 1 h and then atroom temperature for 18 h. The reaction mixture was filtered throughCelite and the filter pad was washed with water and methanol and thefiltrate was evaporated. The residue was acidified to pH 2 withhydrochloric acid (2 M) and the mixture was extracted with ethyl acetateand chloroform. The organic extracts were combined and washed with asmall volume of brine. The organic solution was filtered through ahydrophobic frit and the filtrate was evaporated to afford crude5,5-dimethyl-[1,4]dioxane-2-carboxylic acid (694 mg, 4.5 mmol) as a waxysolid. LCMS (m/z) 159.1 [M−H], Tr=0.88 min. A solution of5,5-dimethyl-[1,4]dioxane-2-carboxylic acid (750 mg, 4.7 mmol), oxalylchloride (600 mg, 0.4 mL, 4.7 mmol) and N,N-dimethylformamide (2 drops)in dichloromethane (15 mL) was stirred at room temperature undernitrogen for 2 h. A solution of benzyl alcohol (508 mg, 0.5 mL, 4.7mmol) in dichloromethane (2 mL) was added and the reaction mixture wasstirred at room temperature for 20 h. Additional benzyl alcohol (0.5 mL,4.7 mmol) was added and the reaction mixture was stirred at roomtemperature for 18 h. The solvent was evaporated and the residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 9:1 to 1:1 to afford the title compound (870mg, 74%) as an oil. ¹H NMR (300 MHz, CDCl₃) δ1.19 (s, 3H), 1.32 (s, 3H),3.43 (d, J=11.4 Hz, 1H), 3.72 (d, J=11.4 Hz, 1H), 3.88-3.92 (m, 2H),4.21 (dd, J=8.2, 4.7 Hz, 1H), 5.24 (s, 2H), 7.34-7.39 (m, 5H). LCMS(m/z) 273.1 [M+Na], Tr=2.41 min.

Compound 100d.2-(1-Hydroxy-ethyl)-5,5-dimethyl-[1,4]dioxane-2-carboxylic acid benzylester

A solution of N,N-diisopropylamine (530 mg, 0.73 mL, 5.25 mmol) inanhydrous tetrahydrofuran (4 mL) was stirred at −78° C. under nitrogen.n-Butyl lithium (2.1 mL, 5.25 mmol, 2.5 M solution in hexane) was addeddropwise and the reaction mixture was stirred at −78° C. for 30 min. Asolution of 5,5-dimethyl-[1,4]dioxane-2-carboxylic acid benzyl ester(870 mg, 3.5 mmol) in tetrahydrofuran (8 mL) was added and the reactionmixture was stirred at −78° C. for 20 min. Acetaldehyde (462 mg, 0.6 mL,10.5 mmol) was added in one portion. The cooling bath was removed andthe reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was cooled to 0° C. and ice-cold hydrochloric acid (2M) was added to acidify the reaction mixture to pH 2. Sodium chloridewas added to saturate the aqueous phase and the mixture was extractedwith diethyl ether. The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 6:1 to 0:1followed by silica gel chromatography using iso-hexanes/ethyl acetate3:1 to 0:1 to the title compound (400 mg, 40%) as an oil and as a 2:1mixture of diastereoisomers. LCMS (m/z) 295.1 [M+H], Tr=2.11 min.

Compound 100e. 5,5-Dimethyl-2-vinyl-[1,4]dioxane-2-carboxylic acidbenzyl ester

A suspension of sodium hydride (80 mg, 2 mmol, 60% dispersion in mineraloil) in anhydrous tetrahydrofuran (3 mL) was stirred at −78° C. undernitrogen. A solution of2-(1-hydroxy-ethyl)-5,5-dimethyl-[1,4]dioxane-2-carboxylic acid benzylester (400 mg, 1.36 mmol) in tetrahydrofuran (3 mL) was added and thereaction mixture was stirred at −78° C. for 15 min. A solution ofN-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (1.06 g, 2.7mmol) in tetrahydrofuran (5 mL) was added and the reaction mixture wasstirred at −78° C. for 30 min. The reaction mixture was warmed to roomtemperature and then stirred at room temperature for 5 h. The reactionmixture was cooled to 5° C., saturated ammonium chloride solution wascautiously added and the mixture was extracted with ethyl acetate. Theorganic extracts were combined and washed with brine. The organicsolution was filtered through a hydrophobic frit and the filtrate wasevaporated. The residue was purified by silica gel chromatography usinga gradient of iso-hexanes/ethyl acetate 9:1 to 4:1 to afford crude5,5-dimethyl-2-(1-trifluoromethanesulfonyloxy-ethyl)-[1,4]dioxane-2-carboxylicacid benzyl ester (833 mg, 1.3 mmol) as a yellow oil which darkened uponstanding and as a 2:1 mixture of diastereoisomers which was usedimmediately in the next step. LCMS (m/z) 449.0 [M+H], Tr=3.29 min. Asolution of5,5-dimethyl-2-(1-trifluoromethanesulfonyloxy-ethyl)-[1,4]dioxane-2-carboxylicacid benzyl ester (833 mg, 1.3 mmol) in dichloromethane (5 mL) wasstirred at room temperature under nitrogen. 1,8-Diazabicycloundec-7-ene(790 mg, 0.8 mL, 5.2 mmol) was added and the reaction mixture wasstirred at room temperature for 72 h. The reaction mixture was cooled to0° C. and acidified to pH 2 with ice cold hydrochloric acid (2 M). Themixture was extracted with dichloromethane. The organic extracts werecombined and washed with brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography using a gradient ofiso-hexanes/diethyl ether 9:1 to 3:1 to afford the title compound (189mg, 52% over two steps) as a colorless oil. ¹H NMR (300 MHz, CDCl₃):δ1.13 (s, 3H), 1.32 (s, 3H), 3.53 (d, J=11.8 Hz, 1H), 3.66 (d, J=11.8Hz, 1H), 3.72 (d, J=12.1 Hz, 1H), 4.24 (d, J=12.1 Hz, 1H), 5.19-5.35 (m,3H), 5.51 (dd, J=17.4, 0.9 Hz, 1H), 5.80 (dd, J=17.4, 10.7 Hz, 1H),7.35-7.40 (m, 5H). LCMS (m/z) 277.1 [M+H], Tr=2.81 min.

Compound 100f. 5,5-Dimethyl-2-vinyl-[1,4]dioxane-2-carboxylic acid

To a stirred solution of 5,5-dimethyl-2-vinyl-[1,4]dioxane-2-carboxylicacid benzyl ester (189 mg, 0.684 mmol) in tetrahydrofuran (7 mL) at 0°C. was added a solution of lithium hydroxide hydrate (34 mg, 0.821 mmol)in water (1.6 mL). The reaction mixture was stirred at 0° C. for 30 minand allowed to warm to ambient temperature and stirred for 19 h. Themixture was diluted with water and washed with ethyl acetate (2×). Theaqueous layer was acidified to pH 5 with hydrochloric acid (2 M) andextracted with ethyl acetate (2×). The organic extracts were dried overanhydrous magnesium sulfate, filtered and evaporated to give the titlecompound (69 mg, 54%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃): δ1.19(s, 3H), 1.32 (s, 3H), 3.58 (d, J=11.8 Hz, 1H), 3.72 (d, J=11.8 Hz, 1H),3.75 (d, J=12.3 Hz, 1H), 4.21 (d, J=12.3 Hz, 1H), 5.39 (dd, J=10.7, 0.9Hz, 1H), 5.60 (dd, J=17.4, 0.9 Hz, 1H), 5.84 (dd, J=17.4, 10.7 Hz, 1H).LCMS (m/z) 185.2 [M−H], Tr=1.32 min.

Compound 100 g.2-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-5,5-dimethyl[1,4]dioxane-2-carboxylicacid

A mixture of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (198 mg, 0.371 mmol),5,5-dimethyl-2-vinyl-[1,4]dioxane-2-carboxylic acid (69 mg, 0.371 mmol),triethylamine (112 mg, 155 μL, 1.11 mmol) and tri(o-tolyl)phosphine (23mg, 0.074 mmol) in 1,4-dioxane (7.5 mL) was degassed with nitrogen for 5min and then warmed to 50° C. under nitrogen with stirring.Tris(dibenzylideneacetone)dipalladium(0) (34 mg, 0.037 mmol) was addedand the mixture was heated to 80° C. for 40 min and then allowed to coolto room temperature. The suspension was filtered and evaporated to givecrude title compound (0.371 mmol) which was used directly in the nextstage. LCMS (m/z) 640.3 [M+H], Tr=1.94 min.

Compounds 100 and 101

To a stirred mixture of powdered 4 Å molecular sieves (˜1 g),2-methyl-6-nitrobenzoic anhydride (255 mg, 0.742 mmol) and4-(dimethylamino)-pyridine (181 mg, 1.48 mmol) in dichloromethane (111mL), under nitrogen, was added a solution of the2-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-5,5-dimethyl[1,4]dioxane-2-carboxylicacid (0.371 mmol) in dichloromethane (5 mL) over 2.5 h via syringe pump.The flask originally containing the crude acid was washed withdichloromethane (1 mL) and this solution was added to the reactionmixture over 30 min. After the end of addition, the reaction mixture wasstirred for 30 min, filtered through Celite and washed successively withice cold saturated ammonium chloride solution (2×) and ice coldsaturated sodium bicarbonate solution (2×), passed through a hydrophobicfrit and evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/acetone 1:0 to 1:1followed by reverse phase preparative HPLC on an Agilent Eclipse XDB/C187 micron, 250×21.2 mm column (flow rate 20 mL/min) usingacetonitrile/water 9:11 to give the first eluting more polardiastereoisomer (2.9 mg, 1% over 2 steps) as a white solid and thesecond eluting less polar diastereoisomer (6.6 mg, 3% over 2 steps) as awhite solid.

First eluting diastereoisomer. Compound 100 ¹H NMR (300 MHz, CD₃OD):δ1.07 (d, J=6.7 Hz, 3H), 1.12 (s, 3H), 1.13 (d, J=6.9 Hz, 3H), 1.43 (s,3H), 1.52-1.78 (m, 2H), 1.57 (d, J=7.5 Hz, 3H), 1.59 (d, J=6.7 Hz, 3H),1.93-2.02 (m, 1H), 2.18-2.39 (m, 2H), 2.68-2.80 (m, 1H), 3.55-3.64 (m,2H), 3.73 (d, J=11.8 Hz, 1H), 3.92 (d, J=12.3 Hz, 1H), 4.39 (d, J=12.3Hz, 1H), 4.41-4.50 (m, 1H), 5.11 (q, J=6.7 Hz, 1H), 5.39 (d, J=8.5 Hz,1H), 5.83 (q, J=6.9 Hz, 1H), 6.31 (d, J=16.5 Hz, 1H), 6.98 (d, J=16.5Hz, 1H), 7.49 (d, J=8.7 Hz, 1H), 7.65 (dd, J=8.5, 1.4 Hz, 1H), 7.85 (s,1H), 7.89 (d, J=8.5 Hz, 1H), 8.28 (d, J=8.5 Hz, 1H). LCMS (m/z) 622.2[M+H], Tr=2.48 min.

Second eluting diastereoisomer. Compound 101 ¹H NMR (300 MHz, CD₃OD):δ1.08 (d, J=6.7 Hz, 3H), 1.09 (d, J=6.5 Hz, 3H), 1.13 (s, 3H), 1.43 (s,3H), 1.52-1.76 (m, 2H), 1.59 (d, J=6.7 Hz, 3H), 1.64 (d, J=7.4 Hz, 3H),1.89-1.99 (m, 1H), 2.19-2.28 (m, 2H), 2.62-2.73 (m, 1H), 3.55-3.71 (m,3H), 3.87 (d, J=12.0 Hz, 1H), 4.30 (d, J=11.8 Hz, 1H), 4.37-4.47 (m,1H), 5.09 (q, J=6.7 Hz, 1H), 5.21 (d, J=8.9 Hz, 1H), 5.79 (q, J=7.2 Hz,1H), 6.14 (d, J=16.3 Hz, 1H), 6.89 (d, J=16.3 Hz, 1H), 7.47 (d, J=8.5Hz, 1H), 7.65 (dd, J=8.5, 1.6 Hz, 1H), 7.86 (d, J=8.2 Hz, 1H), 7.88 (s,1H), 8.26 (d, J=8.5 Hz, 1H). LCMS (m/z) 622.3 [M+H], Tr=2.47 min.

Examples 102, 103, 104 and 105 Compounds 102, 103, 104 and 105

Compound 102a. 3-Triisopropylsilanyloxy-cyclopentanecarboxylic acidethyl ester

A solution of 3-hydroxy-cyclopentanecarboxylic acid ethyl ester(prepared as described in PCT Int. Appl. 2008131946, 3.180 g, 20.1 mmol)in dichloromethane (100 mL) was subsequently treated with 2,6-lutidine(4.7 mL, 40.2 mmol) and triisopropylsilyl trifluoromethanesulfonate (8.1mL, 30.15 mmol). After stirring for 3.5 h, the reaction was quenchedwith pH 4 buffer. The aqueous layer was extracted with dichloromethane.The organics were combined, filtered through a phase separator and thevolatiles were removed in vacuo. The residue was purified by silica gelchromatography using a 100 g Isolute cartridge eluted with a continuousgradient of iso-hexanes/diethyl ether 1:0 to 9:1 to afford the titlecompound (6.15 g, 97%) as a light yellow oil and as a 3.5:1 mixture ofdiastereoisomers. ¹H NMR (300 MHz, CDCl₃): δ1.04-1.08 (m, 27H), 1.27 (t,J=7.1 Hz, 4H), 1.70-2.00 (m, 5.2H), 2.04-2.21 (m, 2.6H), 2.66-2.79 (m,1H), 2.97-3.10 (m, 0.3H), 4.14 (q, J=7.1 Hz, 2.6H), 4.29-4.39 (m, 1H),4.43-4.50 (m, 0.3H).

Compound 102b.1-(1-Hydroxy-ethyl)-3-triisopropylsilanyloxy-cyclopentanecarboxylic acidethyl ester

A cooled (−78° C.) solution of N,N-diisopropylamine (4.4 mL, 31.285mmol) in anhydrous tetrahydrofuran (60 mL) was treated withn-butyllithium (2.5 M in hexanes, 11.7 mL, 29.329 mmol). After stirringat −78° C. for 30 min, the reaction mixture was treated with a solutionof 3-triisopropylsilanyloxy-cyclopentanecarboxylic acid ethyl ester(6.15 g, 19.553 mmol) in anhydrous tetrahydrofuran (20 mL). Afterstirring at −78° C. for 40 min, the reaction mixture was treated withneat acetaldehyde (3.3 mL, 58.659 mmol). The cooling bath was removedand the reaction was stirred at room temperature for 50 min thenquenched at 0° C. with a saturated solution of ammonium chloride (100mL). The aqueous layer was extracted with dichloromethane (2×). Theorganics were combined, filtered through a phase separator and thevolatiles were removed in vacuo. The residue was purified by silica gelchromatography using a 100 g Isolute cartridge eluted with a continuousgradient of iso-hexanes/ethyl acetate 1:0 to 4:1 to afford the titlecompound (6.61 g, 94%) as a light yellow oil and as a complex mixture ofdiastereoisomers.

Compound 102c. 3-Triisopropylsilanyloxy-1-vinyl-cyclopentanecarboxylicacid

A cooled (−78° C.) suspension of sodium hydride (1.474 g, 36.868 mmol,60% dispersed in mineral oil) in anhydrous tetrahydrofuran (60 mL) wastreated with a solution of1-(1-hydroxy-ethyl)-3-triisopropylsilanyloxy-cyclopentanecarboxylic acidethyl ester (6.61 g, 18.434 mmol). After stirring at −78° C. for 10 min,the reaction mixture was treated with a solution ofN-(5-chloro-2-pyridyl)bis(trifluoromethanesulfonimide) (13.2 g, 33.615mmol) in anhydrous tetrahydrofuran (20 mL). The cooling bath was removedand the reaction was allowed to stir at room temperature for 24 h. Moresodium hydride (1.474 g, 36.868 mmol, 60% dispersed in mineral oil) wasadded. After stirring at room temperature for 5 h, more sodium hydride(1.474 g, 36.868 mmol, 60% dispersed in mineral oil) was added. Afterstirring at room temperature for 17.5 h, the reaction was quenched at 0°C. with a saturated solution of ammonium chloride (100 mL). The aqueouslayer was extracted with dichloromethane (2×). The aqueous layer wasacidified to pH 2 with hydrochloric acid (2 M) and extracted withdichloromethane. The organics were combined, filtered through a phaseseparator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 100 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/ethyl acetate 1:0 to4:1 followed by silica gel chromatography using a 100 g Isolutecartridge eluted with a continuous gradient of iso-hexanes/diethyl ether1:0 to 4:1 to afford the title compound (876.9 mg, 15%) as a lightyellow oil and as a 3:1 mixture of diastereoisomers. LCMS (m/z) 311.2[M−H], Tr=4.05 min.

Compound 102d.1-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-Cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-3-triisopropylsilanyloxy-cyclopentanecarboxylicacid

A solution of(S)-1-[(S)-2-((S)-2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (795.0 mg, 1.493 mmol),3-triisopropylsilanyloxy-1-vinyl-cyclopentanecarboxylic acid (466.6 mg,1.493 mmol), tri(o-tolyl)phosphine (91.0 mg, 0.299 mmol) andtriethylamine (0.63 mL, 4.479 mmol) in 1,4-dioxane (20 mL) wasdeoxygenated by bubbling through nitrogen for 5 min. The reactionmixture was then treated with tris(dibenzylideneacetone)dipalladium(0)(136.7 mg, 0.149 mmol). After stirring at 100° C. for 40 min, thereaction was cooled to room temperature and filtered through Celite. Thesolid was rinsed with ethyl acetate and the volatiles were removed invacuo to provide the crude title compound (1.493 mmol) as an orangefoam. LCMS (m/z) 764.4 [M−H], Tr=3.53 min.

Compound 102e

A solution of crude1-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-cyclopropyl-2-hydroxy-acetylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-3-triisopropylsilanyloxy-cyclopentanecarboxylicacid (1.493 mmol) in dry dichloromethane (20 mL) was added via syringepump to a solution of 2-methyl-6-nitrobenzoic anhydride (771.0 mg, 2.239mmol), 4-dimethylaminopyridine (547.1 mg, 4.479 mmol) in drydichloromethane (500 mL) containing 4 Å molecular sieves over 4.5 h.After the end of the addition, the reaction was stirred at roomtemperature for 2 h then filtered and the volatiles were partiallyremoved in vacuo. The organics were washed with hydrochloric acid (0.5M, 40 mL). The acidic aqueous layer was extracted with dichloromethane.The combined organics were washed with a saturated solution of sodiumbicarbonate. The basic aqueous layer was extracted with dichloromethane.The combined organics were filtered on a phase separator. The volatileswere removed in vacuo and the residue was purified by silica gelchromatography using a 50 g Isolute cartridge eluted with a continuousgradient of iso-hexanes/acetone 1:0 to 3:2 to afford the title compound(459.6 mg, 41%) as a white solid and as a complex mixture ofdiastereoisomers. LCMS (m/z) 746.5 [M+H], Tr=4.35 min.

Compounds 102, 103, 104 and 105

A solution of compound 102e (459.6 mg, 0.616 mmol) inacetonitrile/tetrahydrofuran (20 mL, 9:1) in a 250 mL round-bottomTeflon flask was treated with hydrofluoric acid (48 wt % in water, 1.3mL, 30.8 mmol). After stirring at room temperature for 17 h, morehydrofluoric acid (48 wt % in water, 1.3 mL, 30.8 mmol) was added. Afterstirring at room temperature for 7 h, the reaction mixture was slowlypoured over a saturated solution of sodium bicarbonate (100 mL). Afterthe bubbling has stopped, the aqueous layer was extracted withdichloromethane (2×). The organics were combined, filtered through aphase separator and the volatiles were removed in vacuo. The residue waspurified by silica gel chromatography using a 50 g Isolute cartridgeeluted with a continuous gradient of iso-hexanes/acetone 1:0 to 2:3. Themixture was further purified by reverse phase preparative HPLC elutingwith a continuous gradient of water (modified with 0.1% trifluoroaceticacid)/acetonitrile (modified with 0.1% trifluoroacetic acid) 9:1 to 3:2collecting 2 mL fractions to afford, after evaporation of volatiles fourdifferent isomers as trifluoroacetate salts.

Compound 102 (41.2 mg, 9.5%) as a light yellow solid. ¹H NMR (300 MHz,CD₃OD): δ 0.46-0.56 (m, 1H), 0.58-0.72 (m, 3H), 1.23-1.36 (m, 1H),1.56-1.67 (m, 7H), 1.69-1.85 (m, 3H), 1.90-2.00 (m, 1H), 2.01-2.12 (m,1H), 2.13-2.24 (m, 2H), 2.46-2.54 (m, 1H), 2.62-2.79 (m, 2H), 3.58-3.66(m, 1H), 4.37-4.47 (m, 2H), 4.94 (d, J=8.7 Hz, 1H), 5.12 (q, J=6.7 Hz,1H), 5.78-5.91 (m, 1H), 6.50 (app d, J=3.1 Hz, 2H), 7.57 (d, J=8.5 Hz,1H), 7.73 (d, J=8.5 Hz, 1H), 7.89 (s, 1H), 7.93 (d, J=8.5 Hz, 1H), 8.43(d, J=8.5 Hz, 1H), 8.66 (d, J=7.1 Hz, 1H). LCMS (m/z) 590.2 [M+H],Tr=1.97 min.

Compound 103 (31.2 mg, 7.2%) as a light yellow gum. ¹H NMR (300 MHz,CD₃OD): δ 0.47-0.57 (m, 1H), 0.60-0.74 (m, 3H), 1.25-1.38 (m, 1H),1.54-1.79 (m, 9H), 1.91-2.25 (m, 5H), 2.48-2.90 (m, 3H), 3.58-3.66 (m,1H), 4.37-4.47 (m, 2H), 5.13 (q, J=6.7 Hz, 1H), 5.78-5.91 (m, 1H), 6.48,6.55 (ABq, J_(AB)=16.3 Hz, 2H), 7.57 (d, J=8.5 Hz, 1H), 7.73 (d, J=8.5Hz, 1H), 7.83-7.96 (m, 2H), 8.43 (d, J=8.5 Hz, 1H), 8.66 (d, J=7.8 Hz,1H). LCMS (m/z) 590.2 [M+H], Tr=1.88 min.

Compound 104 (13.5 mg, 3.1%) as a light yellow solid. ¹H NMR (300 MHz,CD₃OD): δ 0.44-0.57 (m, 1H), 0.61-0.71 (m, 3H), 1.28-1.40 (m, 1H),1.54-1.81 (m, 9H), 1.91-2.13 (m, 4H), 2.17-2.29 (m, 1H), 2.43-2.59 (m,1H), 2.66-2.78 (m, 2H), 3.58-3.66 (m, 1H), 4.30-4.38 (m, 1H), 4.40-4.48(m, 1H), 5.05-5.15 (m, 1H), 5.78-5.91 (m, 1H), 6.47, 6.54 (ABq,J_(AB)=15.8 Hz, 2H), 7.50 (d, J=8.5 Hz, 1H), 7.63-7.74 (m, 1H), 7.80 (s,1H), 7.88 (d, J=8.5 Hz, 1H), 8.32 (d, J=8.5 Hz, 1H), 8.69-8.76 (m, 1H).LCMS (m/z) 590.2 [M+H], Tr=1.93 min.

Compound 105 (6.0 mg, 1.4%) as a light yellow gum. ¹H NMR (300 MHz,CD₃OD): δ 0.45-0.55 (m, 1H), 0.59-0.72 (m, 3H), 1.27-1.44 (m, 1H),1.54-1.84 (m, 9H), 1.90-2.00 (m, 2H), 2.01-2.27 (m, 4H), 2.45-2.54 (m,1H), 2.62-2.81 (m, 2H), 3.57-3.66 (m, 1H), 4.37-4.47 (m, 2H), 4.96 (d,J=8.9 Hz, 1H), 5.11 (q, J=6.9 Hz, 1H), 5.78-5.90 (m, 1H), 6.39-6.62 (m,2H), 7.52 (d, J=8.5 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 7.80-7.92 (m, 2H),8.35 (d, J=8.5 Hz, 1H), 8.66-8.74 (m, 1H). LCMS (m/z) 590.2 [M+H],Tr=1.97 min.

Example 106 Compound 106

Compound 106a. 4-(1-Hydroxy-ethyl)-tetrahydro-thiopyran-4-carboxylicacid methyl ester

A solution of N,N-diisopropylamine (1.51 g, 2.1 mL, 15 mmol) inanhydrous tetrahydrofuran (10 mL) was stirred at −78° C. under nitrogen.n-Butyl lithium (6 mL, 15 mmol, 2.5 M solution in hexane) was addeddropwise and the reaction mixture was stirred at −78° C. for 30 min. Asolution of tetrahydro-thiopyran-4-carboxylic acid methyl ester (1.60 g,10 mmol) in tetrahydrofuran (4 mL) was added and the reaction mixturewas stirred at −78° C. for 20 min. Acetaldehyde (1.32 g, 1.7 mL, 30mmol) was then added in one portion. The cooling bath was removed andthe reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was cooled to 0° C. and ice-cold hydrochloric acid (2M) was added to acidify the reaction mixture to pH 2. Sodium chloridewas added to saturate the aqueous phase and the mixture was extractedwith diethyl ether. The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of pentane/diethyl ether 3:1 to 1:3 toafford the title compound (1.77 g, 87%) as a yellow oil. ¹H NMR (300MHz, d₆-DMSO): δ0.96 (d, J=6.5 Hz, 3H), 1.46-1.73 (m, 2H), 2.12-2.28 (m,2H), 2.43-2.57 (m, 4H), 3.50-3.58 (m, 1H), 3.63 (s, 3H), 4.85 (d, J=5.8Hz, 1H). LCMS (m/z) 227.1 [M+Na], Tr=1.53 min.

Compound 106b.4-(1-Hydroxy-ethyl)-1,1-dioxo-hexahydro-thiopyran-4-carboxylic acidmethyl ester

A solution of 4-(1-hydroxy-ethyl)-tetrahydro-thiopyran-4-carboxylic acidmethyl ester (408 mg, 2 mmol) in dichloromethane (10 mL) was stirred at0° C. Meta-chloroperbenzoic acid (70% pure containing 20% water and 10%meta-chlorobenzoic acid, 830 mg, 4.8 mmol) was added and the reactionmixture was stirred at 0° C. for 3 h. Additional meta-chloroperbenzoicacid (70% pure containing 20% water and 10% meta-chlorobenzoic acid, 830mg, 4.8 mmol) was added and the reaction mixture was stirred at 0° C.for 3 h. Sodium thiosulfate solution and saturated sodium hydrogencarbonate solution were added and the mixture was extracted withdichloromethane. The organic extracts were combined and washed withbrine. The organic solution was filtered through a hydrophobic frit andthe filtrate was evaporated. The residue was purified by silica gelchromatography using a gradient of iso-hexanes/ethyl acetate 1:1 to 0:1to afford the title compound (281 mg, 60%) as a white solid. ¹H NMR (300MHz, d₆-DMSO) 60.99 (d, J=6.5 Hz, 3H), 1.90-2.30 (m, 4H), 2.92-3.11 (m,4H), 3.64-3.71 (m, 4H), 5.12 (d, J=5.8 Hz, 1H). LCMS (m/z) 237.1 [M+H],Tr=0.76 min.

Compound 106c. 1,1-Dioxo-4-vinyl-hexahydro-thiopyran-4-carboxylic acidmethyl ester

A solution of4-(1-hydroxy-ethyl)-1,1-dioxo-hexahydro-thiopyran-4-carboxylic acidmethyl ester (281 mg, 1.2 mmol) in dichloromethane (10 mL) was stirredat 0° C. under nitrogen. Pyridine (284 mg, 0.3 mL, 3.6 mmol) was addedand then trifluoromethanesulfonic anhydride (677 mg, 0.4 mL, 2.4 mmol)was added drop wise and the reaction mixture was stirred at 0° C. for 30min. Saturated sodium hydrogen carbonate solution was added and theorganic layer was separated and washed with brine and filtered through ahydrophobic frit. The filtrate was evaporated to a volume of ˜5 mL andthe solution of crude1,1-dioxo-4-(1-trifluoromethanesulfonyloxy-ethyl)-hexahydro-thiopyran-4-carboxylicacid methyl ester (1.2 mmol) was used immediately in the next step. LCMS(m/z) 368.9 [M+H], Tr=2.27 min. A solution of crude1,1-dioxo-4-(1-trifluoromethanesulfonyloxy-ethyl)-hexahydro-thiopyran-4-carboxylicacid methyl ester (1.2 mmol) in dichloromethane (5 mL) was stirred atroom temperature under nitrogen. 1,8-Diazabicycloundec-7-ene (0.6 mL,3.6 mmol) was added and the reaction mixture was stirred at roomtemperature for 45 min. The reaction mixture was cooled to 0° C. andacidified to pH 2 with ice-cold hydrochloric acid (2 M). The mixture wasextracted with dichloromethane. The organic extracts were combined,washed with water and brine. The organic solution was filtered through ahydrophobic frit and the filtrate was evaporated. The residue waspurified by silica gel chromatography using a gradient ofiso-hexanes/ethyl acetate 4:1 to 1:1 to afford the title compound (182mg, 70% over two steps) as a white solid. ¹H NMR (300 MHz, CDCl₃)δ2.25-2.34 (m, 2H), 2.59-2.66 (m, 2H), 3.05-3.13 (m, 4H), 3.79 (s, 3H),5.23 (d, J=17.6 Hz, 1H), 5.31 (d, J=10.7 Hz, 1H), 5.82 (dd, J=17.6, 10.7Hz, 1H). LCMS (m/z) 219.1 [M+H], Tr=1.33 min.

Compound 106d. 1,1-Dioxo-4-vinyl-hexahydro-thiopyran-4-carboxylic acid

A solution of 1,1-dioxo-4-vinyl-hexahydro-thiopyran-4-carboxylic acidmethyl ester (174 mg, 0.8 mmol) in tetrahydrofuran (4 mL) and methanol(0.5 mL) was stirred at room temperature under nitrogen. A solution oflithium hydroxide monohydrate (134 mg, 3.2 mmol) in water (1 mL) wasadded and the reaction mixture was stirred at room temperature for 1 h.The reaction mixture was cooled to 0° C. and acidified to pH 2 withhydrochloric acid (2 M). Sodium chloride was added and the mixture wasextracted with ethyl acetate. The organic extracts were combined andwashed with brine. The organic solution was filtered through ahydrophobic frit and the filtrate was evaporated to afford the titlecompound (163 mg, 100%) as a white solid. ¹H NMR (300 MHz, d₆-DMSO)□□2.03-2.12 (m, 2H), 2.33-2.42 (m, 2H), 2.96-3.13 (m, 4H), 5.23 (d,J=17.6 Hz, 1H), 5.25 (d, J=10.7 Hz, 1H), 5.87 (dd, J=17.6, 10.7 Hz, 1H),12.9-13.1 (br s, 1H). LCMS (m/z) 227.1 [M+Na], Tr=0.96 min.

Compound 106

A stirred solution of1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (187 mg, 0.35 mmol),1,1-dioxo-4-vinyl-hexahydro-thiopyran-4-carboxylic acid (79 mg, 0.38mmol), tri(o-tolyl)phosphine (21 mg, 0.07 mmol) and triethylamine (106mg, 0.15 mL, 1.05 mmol) in 1,4-dioxane (10 mL) was degassed withnitrogen for 15 min. The solution was heated to 50° C. andtris(dibenzylideneacetone)dipalladium(0) (32 mg, 0.035 mmol) was added.The reaction mixture was heated at 80° C. for 30 min. The reactionmixture was cooled to room temperature and the mixture was filteredthrough a hydrophobic frit and the filtrate was evaporated to affordcrude4-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-1,1-dioxo-hexahydro-thiopyran-4-carboxylicacid (0.35 mmol) as an orange oil. LCMS (m/z) 658.2 [M+H], Tr=1.57 min.A solution of 2-methyl-6-nitrobenzoic anhydride (241 mg, 0.7 mmol) and4-(dimethylamino)-pyridine (170 mg, 1.4 mmol) in dichloromethane (200mL) containing 4 Å molecular sieves (200 mg) was stirred at roomtemperature under nitrogen. A solution of crude4-((E)-2-{2-[(R)-1-({(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carbonyl}-amino)-ethyl]-quinolin-7-yl}-vinyl)-1,1-dioxo-hexahydro-thiopyran-4-carboxylicacid (0.35 mmol) in dichloromethane (5 mL) was added dropwise over 4 hand the reaction mixture was stirred at room temperature for 4 h. Thereaction mixture was filtered through Celite and the solution wasevaporated to give a volume of ˜30 mL. The solution was washed withice-cold saturated ammonium chloride solution, ice-cold saturated sodiumbicarbonate solution and brine. The organic solution was filteredthrough a hydrophobic frit and the filtrate was evaporated. The residuewas purified by silica gel chromatography eluting with a gradient ofiso-hexanes/ethyl acetate/methanol 1:1:0 to 0:1:0 to 0:9:1 followed byreverse phase preparative HPLC eluting with a gradient of acetonitrile(containing 0.1% trifluoroacetic acid)/water (containing 0.1%trifluoroacetic acid) 9:1 to 1:1. Fractions containing the product werecombined and evaporated. The residue was partitioned betweendichloromethane and saturated sodium hydrogen carbonate solution. Theorganic layer was separated and washed with brine. The organic solutionwas filtered through a hydrophobic frit and the filtrate was evaporatedto afford the title compound (14 mg, 7% over two steps) as a whitesolid. ¹H NMR (300 MHz, CD₃OD): δ1.05 (d, J=6.7 Hz, 3H), 1.09 (d, J=6.7Hz, 3H), 1.59 (d, J=6.7 Hz, 3H), 1.62 (d, J=7.1 Hz, 3H), 1.63-1.71 (m,2H), 1.92-1.97 (m, 1H), 2.17-3.25 (m, 11H), 3.55-3.65 (m, 1H), 4.40-4.46(m, 1H), 5.09 (q, J=6.7 Hz, 1H), 5.33 (d, J=8.3 Hz, 1H), 5.85 (q, J=7.1Hz, 1H), 6.36 (d, J=16.3 Hz, 1H), 6.73 (d, J=16.3 Hz, 1H), 7.47 (d,J=8.5 Hz, 1H), 7.68 (dd, J=8.5, 1.6 Hz, 1H), 7.85 (br s, 1H), 7.87 (d,J=8.5 Hz, 1H), 8.28 (d, J=8.5 Hz, 1H). LCMS (m/z) 640.2 [M+H], Tr=2.09min.

Example 107 Compound 107

Compound 107a. 2,2-Bis-fluoromethyl-5-vinyl-[1,3]dioxane-5-carboxylicacid ethyl ester

A vial was loaded with 2,2-bis-hydroxymethyl-but-3-enoic acid ethylester (0.88 g, 5.05 mmol), 1,3-difluoroacetone (0.95 g, 10.1 mmol) andpara-toluene-4-sulfonic acid monohydrate (0.96 g, 5.05 mmol). The vialwas sealed and heated in a microwave reactor at 100° C. for 15 min. Theresultant brown oil was partitioned between dichloromethane and aqueoussodium bicarbonate solution. The organic layer was filtered through ahydrophobic frit and evaporated to give a brown oil (0.98 g). The oilwas purified by silica gel chromatography using iso-hexanes/ethylacetate 3:1 to yield the title compound (0.619 g, 74%) as a colorlessoil. ¹H NMR (300 MHz, CDCl₃): δ1.31 (t, J=7.1 Hz, 3H), 3.96 (d, J=12.0Hz, 2H), 4.27 (q, J=7.1 Hz, 2H), 4.34-4.69 (m, 6H), 5.22-5.35 (m, 2H),5.67-5.79 (m, 1H).

Compound 107b. 2,2-Bis-fluoromethyl-5-vinyl-[1,3]dioxane-5-carboxylicacid

A solution of lithium hydroxide monohydrate (311 mg, 7.42 mmol) in water(8 mL) was added to a stirred solution of2,2-bis-fluoromethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid ethyl ester(619 mg, 2.47 mmol) in tetrahydrofuran/methanol (12 mL, 4:1). Thereaction mixture was stirred at room temperature for 1 h and then cooledto 0° C. before acidifying to pH 4 with hydrochloric acid (1 M). Themixture was saturated with sodium chloride and then extracted withdichloromethane (3×10 mL). The extract was filtered through ahydrophobic frit and evaporated to give the title product (390 mg, 71%)as a colorless gum. ¹H NMR (300 MHz, CDCl₃) δ4.00 (d, J=12.0 Hz, 2H),4.35-4.73 (m, 6H), 5.30-5.43 (m, 2H), 5.68-5.81 (m, 1H).

Example 107 Compound 107

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (300 mg, 0.56 mmol),2,2-bis-fluoromethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid (125 mg,0.56 mmol), triethylamine (234 μL, 1.68 mmol) and tri(o-tolyl)phosphine(34 mg, 0.112 mmol) in 1,4-dioxane (12 mL) was purged with nitrogen for10 min before adding tris(dibenzylideneacetone) dipalladium(0) (51 mg,0.056 mmol). The stirred reaction mixture was heated at 60° C. for 1 h.The mixture was cooled to room temperature, filtered through Celite thenevaporated to yield an orange foam. The foam was dissolved indichloromethane (10 mL) and the resulting solution was added over 5 hvia syringe pump to a solution of 2-methyl-6-nitro-benzoic anhydride(482 mg, 1.4 mmol) and 4-(dimethylamino)pyridine (350 mg, 2.8 mmol) indichloromethane (205 mL) containing powdered 3 Å sieves. After the endof addition, the solution was stirred for 20 min, filtered, washed withaqueous sodium bicarbonate solution (150 mL), then filtered through ahydrophobic frit and evaporated to give a brown gum which was purifiedby reverse phase preparative HPLC using a gradient of water/acetonitrile4:1 to 0:1 followed by reverse phase preparative HPLC using water(modified with 0.1% ammonium formate)/acetonitrile 3:2 to afford thetitle compound (39 mg, 10%) as a white solid. ¹H NMR (300 MHz, CD₃OD):δ1.06 (d, J=6.7 Hz, 3H), 1.12 (d, J=6.7 Hz, 3H), 1.49-1.78 (m, 2H), 1.61(d, J=6.7 Hz, 3H), 1.66 (d, J=7.1 Hz, 3H), 1.92-2.02 (m, 1H), 2.13-2.30(m, 2H), 2.64-2.78 (m, 1H), 3.58-3.66 (m, 1H), 4.12-4.25 (m, 3H),4.35-4.92 (m, 6H), 5.07 (q, J=6.7 Hz, 1H), 5.38 (d, J=9.1 Hz, 1H), 5.82(q, J=7.1 Hz, 1H), 6.17 (d, J=16.5 Hz, 1H), 6.65 (d, J=16.5 Hz, 1H),7.48 (d, J=8.5 Hz, 1H), 7.67-7.76 (m, 2H), 7.87 (d, J=8.5 Hz, 1H), 8.27(d, J=8.5 Hz, 1H). LCMS (m/z) 658.2 [M+H], Tr=2.46 min.

Example 108 Compound 108

Compound 108

To a solution of the reactant 74 (7.5 mg, 0.012 mmol) in pyridine (3drops) was added n-butyryl chloride (1 drop) at rt. After 1 h at rt, themixture was diluted with ethyl acetate, washed with 10% citric acidsolution (×1) and saturated NaHCO₃ solution (×1). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 4.6 mg(55%) of the product Compound 108. ¹H NMR (400 MHz, Methanol-d₄): δ8.23(d, J=8.5 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.63 (dd,J=8.7, 1.6 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.60 (d, J=16.3 Hz, 1H),6.29 (d, J=16.4 Hz, 1H), 5.80 (q, J=7.2 Hz, 1H), 5.25 (d, J=8.5 Hz, 1H),5.07 (q, J=6.6 Hz, 1H), 4.99 (s, 1H), 4.41 (d, J=13.2 Hz, 1H), 3.57 (m,1H), 2.75-2.59 (m, 1H), 2.34 (t, J=7.3 Hz, 2H), 2.31-2.06 (m, 3H),2.05-1.75 (m, 6H), 1.75-1.62 (m, 3H), 1.60 (d, J=7.0 Hz, 3H), 1.57 (d,J=7.0 Hz, 3H), 1.28 (s, 3H), 1.06 (d, J=6.8 Hz, 3H), 1.01 (d, J=6.8 Hz,3H), 0.98 (t, J=6.8 Hz, 3H).

LCMS (m/z) 676.4 [M+H], Tr 1.77 min (3 min run).

Example 109 Compound 109

Compound 109

To a solution of the 74 (7.3 mg, 0.012 mmol) in pyridine (3 drops) wasadded propionyl chloride (2 drops) at rt. After 5 min, ethyl acetate (˜1mL) was added and sonicated to make it a suspension. After 20 min, themixture was diluted with ethyl acetate, washed with 10% citric acidsolution (×1) and saturated NaHCO₃ solution (×1). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 6.0 mg(75%) of the product Compound 109. ¹H NMR (400 MHz, Methanol-d₄): δ 8.46(d, J=5.6 Hz, 1H), 8.23 (d, J=8.5 Hz, 1H), 7.83 (d, J=8.5 Hz, 1H), 7.78(s, 1H), 7.63 (dd, J=8.5, 1.7 Hz, 1H), 7.43 (d, J=8.4 Hz, 1H), 6.60 (d,J=16.3 Hz, 1H), 6.29 (d, J=16.3 Hz, 1H), 5.80 (q, J=7.2 Hz, 1H), 5.25(d, J=8.5 Hz, 1H), 5.07 (q, J=6.5 Hz, 1H), 4.98 (s, 1H), 4.47-4.34 (m,1H), 3.57 (dd, J=11.8, 3.0 Hz, 1H), 2.68 (td, J=13.0, 3.1 Hz, 1H), 2.38(q, J=7.6 Hz, 2H), 2.28 (dd, J=24.6, 13.3 Hz, 1H), 2.20-2.07 (m, 1H),2.05-1.74 (m, 5H), 1.74-1.63 (m, 1H), 1.60 (d, J=7.3 Hz, 3H), 1.57 (d,J=6.7 Hz, 3H), 1.37-1.19 (m, 3H), 1.15 (t, J=7.6 Hz, 3H), 1.06 (d, J=6.8Hz, 3H), 1.01 (d, J=6.6 Hz, 3H), 0.94-0.67 (m, 2H).

LCMS (m/z) 662.4 [M+H], Tr 1.65 min (3 min run).

Example 110 Compound 110

Compound 110

To a solution of the reactant 74 (7.1 mg, 0.012 mmol) in pyridine (3drops) was added acetic anhydride (2 drops) at rt. After 21 h, themixture was diluted with ethyl acetate, washed with 10% citric acidsolution (×1) and saturated NaHCO₃ solution (×1). After the aqueousfractions were extracted with ethyl acetate (×1), two organic fractionswere combined, dried (MgSO₄), and concentrated. The residue was purifiedby CombiFlash using hexanes-ethyl acetate as eluents to obtain 6.6 mg(87%) of the product Compound 110. ¹H NMR (400 MHz, Methanol-d₄): δ 8.23(d, J=8.5 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 7.78 (s, 1H), 7.62 (dd,J=8.4, 1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H), 6.61 (d, J=16.3 Hz, 1H),6.29 (d, J=16.3 Hz, 1H), 5.80 (q, J=7.2 Hz, 1H), 5.25 (d, J=8.5 Hz, 1H),5.07 (q, J=6.6 Hz, 1H), 4.96 (s, 1H), 4.41 (dd, J=13.5, 4.3 Hz, 1H),3.57 (dd, J=11.8, 2.9 Hz, 1H), 2.68 (td, J=13.0, 3.2 Hz, 1H), 2.39-2.20(m, 2H), 2.20-2.09 (m, 1H), 2.07 (s, 3H), 2.05-1.75 (m, 6H), 1.75-1.62(m, 2H), 1.60 (d, J=7.3 Hz, 3H), 1.57 (d, J=6.7 Hz, 3H), 1.06 (d, J=6.8Hz, 3H), 1.01 (d, J=6.6 Hz, 3H), 0.98-0.69 (m, 2H).

LCMS (m/z) 648.5 [M+H], Tr 1.54 min (3 min run).

Example 111 Compound 111

Compound 111

To a solution of the reactant 75 (7.1 mg, 0.012 mmol) in pyridine (3drops) was added propionyl chloride (2 drops) at rt. After 5 min, ethylacetate (˜1 mL) was added and sonicated to make it a suspension. After20 min, the mixture was diluted with ethyl acetate, washed with 10%citric acid solution (×1) and saturated NaHCO₃ solution (×1). After theaqueous fractions were extracted with ethyl acetate (×1), two organicfractions were combined, dried (MgSO₄), and concentrated. The residuewas purified by CombiFlash using hexanes-ethyl acetate as eluents toobtain 7.3 mg (94%) of the product Compound 111. ¹H NMR (400 MHz,Methanol-d₄): δ 8.23 (d, J=8.5 Hz, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.77 (d,J=1.7 Hz, 1H), 7.61 (dd, J=8.5, 1.7 Hz, 1H), 7.43 (d, J=8.5 Hz, 1H),6.55 (d, J=16.4 Hz, 1H), 6.26 (d, J=16.4 Hz, 1H), 5.80 (q, J=7.2 Hz,1H), 5.28 (d, J=8.6 Hz, 1H), 5.06 (q, J=6.4 Hz, 1H), 4.78 (dt, J=10.8,6.2 Hz, 1H), 4.41 (d, J=13.3 Hz, 1H), 3.58 (dd, J=11.7, 3.1 Hz, 1H),2.68 (td, J=12.9, 3.2 Hz, 1H), 2.58 (d, J=14.1 Hz, 1H), 2.42-2.11 (m,4H), 2.10-1.87 (m, 3H), 1.80-1.63 (m, 3H), 1.61 (d, J=7.2 Hz, 3H), 1.57(d, J=6.8 Hz, 3H), 1.53-1.37 (m, 2H), 1.17-1.06 (m, 6H), 1.03 (d, J=6.6Hz, 3H), 0.96-0.69 (m, 2H).

LCMS (m/z) 662.5 [M+H], Tr 1.66 min (3 min run).

Example 112 Compound 112

Compound 112

To a solution of 75 (7.3 mg, 0.012 mmol) in pyridine (3 drops) was addedacetic anhydride (2 drops) at rt. After 18 h, the mixture was dilutedwith ethyl acetate, washed with 10% citric acid solution (×1) andsaturated NaHCO₃ solution (×1). After the aqueous fractions wereextracted with ethyl acetate (×1), two organic fractions were combined,dried (MgSO₄), and concentrated. The residue was purified by CombiFlashusing hexanes-ethyl acetate as eluents to obtain 5.76 mg (73%) of theproduct Compound 112. ¹H NMR (400 MHz, Methanol-d₄): δ 8.23 (d, J=7.6Hz, 1H), 7.82 (d, J=8.5 Hz, 1H), 7.76 (s, 1H), 7.61 (dd, J=8.5, 1.7 Hz,1H), 7.43 (d, J=8.5 Hz, 1H), 6.55 (d, J=16.3 Hz, 1H), 6.26 (d, J=16.3Hz, 1H), 5.80 (q, J=7.2 Hz, 1H), 5.28 (d, J=8.6 Hz, 1H), 5.06 (m, 1H),4.77 (dt, J=10.6, 6.1 Hz, 1H), 4.48-4.34 (m, 1H), 3.64-3.51 (m, 1H),2.68 (td, J=13.0, 3.3 Hz, 1H), 2.58 (d, J=14.6 Hz, 1H), 2.35 (d, J=12.8Hz, 1H), 2.29-2.21 (m, 1H), 2.17 (ddd, J=13.0, 7.9, 6.4 Hz, 1H), 2.02(s, 3H), 2.10-1.90 (m, 2H), 1.80-1.63 (m, 3H), 1.61 (d, J=7.2 Hz, 3H),1.57 (d, J=6.7 Hz, 3H), 1.54-1.39 (m, 2H), 1.07 (d, J=6.8 Hz, 3H), 1.03(d, J=6.6 Hz, 3H), 0.89 (m, 2H).

LCMS (m/z) 648.4 [M+H], Tr 1.53 min (3 min run).

Example 113 Compound 113

Compound 113a. 2,2-Dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid

A solution of 2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid ethylester (150 mg, 0.7 mmol) in tetrahydrofuran (3 mL) was stirred at 5° C.under nitrogen. A solution of lithium hydroxide monohydrate (59 mg, 1.4mmol) in water (1 mL) was added and the reaction mixture was stirred at5° C. for 30 minutes and then at room temperature for 5 hours. Methanol(0.5 mL) was added to give a clear solution and the reaction mixture wasstirred at room temperature for 22 hours. The solvent was evaporated.Water (2 mL) was added to the residue and the solution was acidified topH 2 with 2 M hydrochloric acid. Brine was added and the mixture wasextracted with ethyl acetate. The organic extracts were combined, washedwith brine. The organic extract was separated, washed with water (×5),and brine. The organic solution was filtered through a hydrophobic fritand the filtrate was evaporated to afford the title compound (117 mg,90%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃) δ1.44 (s, 3H), 1.49 (s,3H), 3.91 (d, J=12.0 Hz, 2H), 4.32 (d, J=12.0 Hz, 2H), 5.30 (d, J=17.8Hz, 1H), 5.35 (d, J=10.7 Hz, 1H), 5.76 (dd, J=17.8, 10.7 Hz, 1H). LCMS(m/z) 185.1 [M−H], Tr=1.26 min.

Compound 113

A solution of(S)-1-[(S)-2-((S)-2-hydroxy-3-methyl-butyrylamino)-propionyl]-hexahydro-pyridazine-3-carboxylicacid [(R)-1-(7-bromo-quinolin-2-yl)-ethyl]-amide (228 mg, 0.426 mmol),2,2-dimethyl-5-vinyl-[1,3]dioxane-5-carboxylic acid (79 mg, 0.426 mmol),triethylamine (178 μL, 1.28 mmol) and tri(o-tolyl)phosphine (26 mg,0.085 mmol) in 1,4-dioxane (9 mL) was purged with nitrogen for 10minutes before adding tris(dibenzylideneacetone)dipalladium(0) (39 mg,0.042 mmol). The stirred reaction mixture was heated at 60° C. for 1 h.The mixture was filtered and evaporated to yield an orange solid. Thesolid was dissolved in dichloromethane (8 mL) and the resulting solutionwas added over 4.25 h, using a syringe pump, to a solution of2-methyl-6-nitro-benzoic anhydride (366 mg, 1.07 mmol) and4-(dimethylamino)pyridine (266 mg, 2.13 mmol) in dichloromethane (156mL) containing powdered 4 Å sieves. After the end of addition, themixture was stirred for 30 minutes, filtered then washed with aqueoussodium bicarbonate solution (75 mL), filtered through a hydrophobic fritand evaporated to give a yellow gum. The gum was purified by reversephase preparative HPLC eluting with water/acetonitrile 3:2 to afford 113(53 mg, 20%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.06 (d, J=6.7Hz, 3H), 1.11 (d, J=6.7 Hz, 3H), 1.35 (s, 3H), 1.55 (s, 3H), 1.58-1.77(m, 2H), 1.60 (d, J=6.7 Hz, 3H), 1.64 (d, J=7.4 Hz, 3H), 1.90-2.00 (m,1H), 2.14-2.29 (m, 2H), 2.65-2.80 (m, 1H), 3.56-3.64 (m, 1H), 4.03-4.16(m, 2H), 4.29-4.37 (m, 1H), 4.39-4.51 (m, 2H), 5.01-5.12 (m, 1H), 5.38(d, J=9.1 Hz, 1H), 5.75-5.87 (m, 1H), 6.17 (d, J=16.7 Hz, 1H), 6.64 (d,J=16.7 Hz, 1H), 7.46 (d, J=8.7 Hz, 1H), 7.66-7.74 (m, 2H), 7.84 (d,J=8.5 Hz, 1H), 8.24 (d, J=8.5 Hz, 1H). LCMS (m/z) 622.3 [M+H], Tr=2.38min.

Example 114 Compound 114

A solution of compound 113 (20 mg, 0.032 mmol) in methanol (1 mL) wasprepared and hydrochloric acid (1 M, 3 mL) was added. The reactionmixture was stirred for 1 h at room temperature and then was neutralisedby addition of saturated sodium bicarbonate solution. Sodium chloridewas added to saturate the solution. The aqueous layer was extracted withdichloromethane, filtered through a hydrophobic frit and evaporated toyield 114 (14 mg, 75%) as a white solid. ¹H NMR (300 MHz, CD₃OD) δ1.01(d, J=6.7 Hz, 3H), 1.04 (d, J=6.7 Hz, 3H), 1.49-1.77 (m, 2H), 1.60 (d,J=6.9 Hz, 3H), 1.65 (d, J=7.4 Hz, 3H), 1.84-1.96 (m, 1H), 2.08-2.28 (m,2H), 2.64-2.77 (m, 1H), 3.56-3.64 (m, 1H), 3.93-4.16 (m, 4H), 4.37-4.46(m, 1H), 5.06 (q, J=6.7 Hz, 1H), 5.32 (d, J=9.1 Hz, 1H), 5.81 (q, J=7.3Hz, 1H), 6.31 (d, J=16.7 Hz, 1H), 6.63 (d, J=16.7 Hz, 1H), 7.45 (d,J=8.5 Hz, 1H), 7.65-7.74 (m, 2H), 7.84 (d, J=8.5 Hz, 1H), 8.24 (d, J=8.5Hz, 1H). LCMS (m/z) 582.5 [M+H], Tr=1.60 min.

Biological Examples Inhibition of Peptidyl-Prolyl Isomerase (PPlase)Activity

The PPlase assay was based on the procedure reported by Janowski et al.(Anal. Biochem. 1997, 252, 299). Assay buffer (1980 L of a solutioncontaining 35 mM HEPES pH 7.8, 50M DTT, and 0.01% NP40) waspre-equilibrated to 10° C. in a quartz cuvette equipped with an overheadstirrer. To this solution was added 10 L of compound in DMSO (finalconcentration: 0.5% DMSO), followed by 5 L of a 2 M stock solution ofcyclophilin A (final concentration: 5 nM). The reaction was initiatedwith the addition of 5 L of 40 mM of the tetrapeptide Succ-AAPF-pNA (100M final concentration) dissolved in a solution of 0.5 M LiCl intrifluoroethanol. Upon the initiation of the reaction, the absorbance ofthe peptide substrate was monitored at 330 nm for five minutes using aBeckman Coulter DU800 spectrophotometer. Progress curves were fit with asingle-exponential decay model to calculate rates. The IC₅₀ values werecalculated with a four-parameter logistic fit using GraphPad Prismsoftware.

Cyclophilin A TR-FRET Competitive Binding Assay

Inhibitor potency was measured using a competitive binding assay with atime-resolved fluorescence resonance energy transfer (TR-FRET) readout.To a reaction buffer consisting of 35 mM HEPES pH 7.8, 100 mM NaCl,0.01% NP-40 (Pierce), 1 mM DTT, and 1% DMSO were added the following: 5nM of cyclophilin A modified at the N-terminus with an 8×histidineaffinity tag (CypA); 150 nM of cyclosporin A modified with a linkerattached to a Cy5 fluorophore (CsA-Cy5); 1 nM Eu-labeled anti-(6×His)antibody (Perkin-Elmer); and test compound at one of variousconcentrations. The total volume of the assay solution was 100 L. Aftera two-hour incubation, the TR-FRET was measured using a Perkin ElmerEnvision plate reader (excitation at 340 nm, emission measured at 590 nmand 665 nm). The signal was calculated as the ratio of the emission at665 nm to that at 590 nm. An IC₅₀ value was calculated using afour-parameter logistic fit.

When tested, certain compounds of this invention were found to inhibitcyclophilin binding as listed in Table 1. The IC₅₀'s (TR-FRET) arepresented as ranges wherein A is ≦100 nM, B is 101 to 1000 nM and C is1001 to 10,000 nM.

Antiviral Activity

The antiviral activity of a compound can be measured using standardscreening protocols: for example, cell-based Flavivirus immunodetectionassay and cell-based Flavivirus cytopathic effect assay as described inU.S. Patent Publication Number US 20130022573, which is herebyincorporated by reference in its entirety.

One aspect of the invention relates to methods of inhibiting viralinfections, comprising the step of treating a sample or subjectsuspected of needing such inhibition with a composition of theinvention. The antiviral activity of a compound of the invention can bemeasured using standard screening protocols that are known.

The anti-HCV activity of the compounds of this invention was tested in ahuman hepatoma Huh-7 cell line harboring a HCV replicon. The assaycomprised the following steps:

Step 1 (Compound Preparation and Serial Dilution):

Serial dilution was performed in 100% DMSO in a 384-well plate. Asolution containing a compound at 225-fold concentration of the startingfinal serial dilution concentration was prepared in 100% DMSO and 15 μLadded to the pre-specified wells in column 3 or 13 of a polypropylene384-well plate. The rest of the 384-well plate was filled with 10 μL100% DMSO except for columns 23 and 24, where 10 μL of 500 μM a HCVprotease inhibitor (ITMN-191) in 100% DMSO was added. The HCV proteaseinhibitor was used a control of 100% inhibition of HCV replication. Theplate was then placed on a Biomek FX Workstation to start the serialdilution. The serial dilution was performed for ten cycles of 3-folddilution from column 3 to 12 or from column 13 to 22.

Step 2 (Cell Culture Plate Preparation and Compound Addition):

To each well of a black polypropylene 384-well plate, 90 μL of cellmedia containing 1600 suspended Huh-7 HCV replicon cells was added witha Biotek uFlow Workstation. A volume of 0.4 μL of the compound solutionwas transferred from the serial dilution plate to the cell culture plateon a Biomek FX Workstation. The DMSO concentration in the final assaycondition was 0.44%. The plates were incubated for 3 days at 37° C. with5% CO₂ and 85% humidity.

Step 3 (Detection of Cytotoxicity and Inhibition of Viral Replication):

a) Assessment of cytotoxicity: The media in the 384-well cell cultureplate was aspirated with a Biotek EL405 plate-washer. A volume of 50 μLof a solution containing 400 nM Calcein AM in 100% PBS was added to eachwell of the plate with a Biotek uFlow Workstation. The plate wasincubated for 30 minutes at room temperature before the fluorescencesignal (emission 490 nm, exitation 520 nm) was measured with a PerkinElmer Envision Plate Reader.

b) Assessment of Inhibition of Viral Replication:

The calcein-PBS solution in the 384-well cell culture plate wasaspirated with a Biotek EL405 plate-washer. A volume of 20 μL ofDual-Glo luciferase buffer (Promega, Dual-Glo Luciferase Assay Reagent,cat. #E298B) was added to each well of the plate with a Biotek uFlowWorkstation. The plate was incubated for 10 minutes at room temperature.A volume of 20 μL of a solution containing 1:100 mixture of Dual-GloStop & Glo substrate (Promega, Dual-Glo Luciferase Assay Reagent, cat.#E313B) and Dual-Glo Stop & Glo buffer (Promega, Dual-Glo LuciferaseAssay Reagent, cat. #E314B) was then added to each well of the platewith a Biotek uFlow Workstation. The plate was incubated at roomtemperature for 10 minutes before the luminescence signal was measuredwith a Perkin Elmer Envision Plate Reader.

Step 4 (Calculation):

The percent cytotoxicity was determined by calcein AM conversion tofluorescent product. The average fluorescent signal from the DMSOcontrol wells were defined as 100% nontoxic. The individual fluorescentsignal from testing compound treated well was divided by the averagesignal from DMSO control wells and then multiplied by 100% to get thepercent viability. The percent anti-HCV replication activity wasdetermined by the luminescence signal from the testing well compared toDMSO controls wells. The background signal was determined by the averageluminescence signal from the HCV protease inhibitor treated wells andwas subtracted from the signal from the testing wells as well as theDMSO control wells. Following 3-fold serial dilutions, the EC₅₀ and CC₅₀values were calculated by fitting % inhibition at each concentration tothe following equation:

% inhibition=100%/[(EC₅₀ /[I])^(b)+1]

where b is Hill's coefficient. See, for reference, Hill, A. V., ThePossible Effects of the Aggregation of the Molecules of Haemoglobin onits Dissociation Curves, J. Physiol. 40: iv-vii. (1910). % inhibitionvalues at a specific concentration, for example 2 μM, can also bederived from the formula above.

When tested, certain compounds of this invention were found to inhibitviral replication as listed in Table 1. A is ≦1 μM, B is 1.1 to 10 μMand C is 10.1 to 100 μM.

When tested, certain compounds of this invention were found to inhibitviral replication as listed in Table 1. The EC₅₀'s are presented as a %inhibition against the 1b replicon.

TABLE 1 TR-FRET Replicon 1b Example IC₅₀ EC₅₀ % inhibition at 1 μM 1 A A97 2 A A 98 3 A A 89 4 A A 100 5 A A 93 6 A A 94 7 A A 94 8 A A 95 9 A A100 10 A A 92 11 A A 96 12 A A 100 13 A A 100 14 A A 100 15 A A 98 16 AB 20 17 B B 39 18 A A 100 19 A A 100 20 B A 88 21 B A 71 22 A A 91 23 AA 97 24 A A 83 25 A A 98 26 B A 68 27 A A 92 28 A A 99 29 A A 99 30 A A84 31 A A 88 32 B B 56 33 A A 97 34 A A 91 35 A A 88 36 A A 99 37 A A 9838 A A 99 39 B B 16 40 A A 99 41 A A 98 42 A A 96 43 A A 99 44 B A 86 45A — — 46 — — — 47 A A 100 48 A A 100 49 A A 100 50 A A 100 51 A A 100 52A A 100 53 A A 99 54 B A 77 55 A A 100 56 A A 99 57 A A 100 58 A — — 59A A 99 60 A A 96 61 B B 46 62 A A 84 63 A A 100 64 A A 100 65 A A 99 66A A 97 67 B A 67 68 A A 93 69 A A 100 70 C A 57 71 A A 96 72 C B −6 73 AA 97 74 A A 100 75 A A 100 76 A A 69 77 B A 56 78 A A 99 79 C B 3 80 A A99 81 A A 100 82 A A 99 83 A A 100 84 C B 10 85 A A 98 86 B A 93 87 A A61 88 A A 100 89 A A 82 90 A A 99 91 A A 100 92 A A 100 93 A A 98 94 A A94 95 A A 88 96 A A 91 97 A A 96 98 A A 90 99 A A 100 100 A A 100 101 AA 98 102 A B 13 103 A A 90 104 A A 56 105 B B 2 106 A A 95 107 A A 100108 A A 99 109 A A 99 110 A A 100 111 A A 99 112 A A 100

The specific pharmacological and biochemical responses observed may varyaccording to and depending on the particular active compound selected orwhether there are present pharmaceutical carriers, as well as the typeof formulation and mode of administration employed, and such expectedvariations or differences in the results are contemplated in accordancewith practice of the present invention.

Although specific embodiments of the present invention are hereinillustrated and described in detail, the invention is not limitedthereto. The above detailed descriptions are provided as exemplary ofthe present invention and should not be construed as constituting anylimitation of the invention.

1. A compound of Formula I:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein: A is a bond,—O—, —S(O)—, —NH—, —N((C₁-C₄)alkyl)- or (C₁-C₂)alkylene; A¹ is(C₁-C₈)alkylene, (C₂-C₅)alkenylene, (C₂-C₅)alkynylene, arylene,heteroarylene, cycloalkylene, heterocycloalkylene, aryl(C₁-C₂)alkylene,heteroaryl(C₁-C₂)alkylene, cycloalkyl(C₁-C₂)alkylene orheterocycloalkyl(C₁-C₂)alkylene, wherein a sp³ carbon atom of A isoptionally replaced by —O—, —S(O)—, —NH— or —N((C₁-C₄)alkyl)-, andwherein a sp³ or sp² carbon atom of A¹ is optionally substituted withone or more groups selected from the group consisting of halo,(C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, aryl, heterocycloalkyl,cycloalkyl, aryl(C₁-C₄)alkyl, cycloalkyl(C₁-C₄)alkyl,heterocycloalkyl(C₁-C₄)alkyl, arylheterocycloalkyl(C₁-C₄)alkyl, —OR⁹,—SR⁹, —S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂; A² is —CH(R⁸)-arylene,—CH(R⁸)-heteroarylene, —CH(R⁸)-heterocycloalkylene,—CH(R⁸)-cycloalkylene, arylene, cycloalkylene, (C₁-C₃)alkylene,(C₂-C₃)alkenylene or (C₂-C₃)alkynylene, wherein A² is optionallysubstituted with one or more substituents selected from the groupconsisting of —OR⁹, —SR⁹, —S(O)R⁹, —S(O)₂R⁹, —N(R⁹)₂, halo,halo(C₁-C₄)alkyl, halo(C₁-C₄)alkoxy, cyano and (C₁-C₈)alkyl; L¹ is—O—C(O)—, —O—CH₂—, —NR¹¹—C(O)—, —NR¹¹—CH₂—, —NH—C(R¹⁰)₂— or —NH—S(O)₂—,wherein each R¹⁰ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₁-C₄)alkenyl or cycloalkyl; and R¹¹ is (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₁-C₄)alkenyl or cycloalkyl; X¹ is a bond, —O—, —NH—, —N((C₁-C₄)alkyl)-or heterocycloalkylene; R¹ and R² are independently H, (C₁-C₄)alkyl,(C₂-C₄)alkenyl, (C₂-C₄)alkynyl, halo, cyano or —C(O)—(C₁-C₄)alkyl; or R¹and R², when taken together with the carbon to which they are bothattached, form —C(═O)—, —C(═S)— or —C(═N(C₁-C₄)alkyl)-; R³ is H or(C₁-C₄)alkyl which is optionally substituted with halo, cyano, hydroxylor (C₁-C₄)alkoxy; R^(4a) and R^(4b) are independently H, (C₁-C₈)alkyl,aryl, aryl(C₁-C₄)alkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl,cycloalkyl or cycloalkyl(C₁-C₄)alkyl, wherein each R^(4a) and R^(4b) isoptionally substituted with one or more substituents selected from thegroup consisting of cyano, —COOH, halo, hydroxyl, amino, (C₁-C₈)alkoxy,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, aryl and heteroaryl; R^(5a)and R^(5b) are independently H, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, (C₁-C₈)alkoxy, aryl, heterocycloalkyl, cycloalkyl,aryl(C₁-C₄)alkyl, cycloalkyl(C₁-C₄)alkyl orheterocycloalkyl(C₁-C₄)alkyl, wherein each R^(5a) and R^(5b) isoptionally substituted with one or more substituents selected from thegroup consisting of —N₃, cyano, —COOH, halo, hydroxyl, amino,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, (C₁-C₈)alkoxy, aryl andheteroaryl, or R^(5a) and R^(5b) together form a spirocycle havingFormula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹; R^(6a) and R^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl,(C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ and —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy, aryl, cycloalkyl, heterocycloalkyl,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹, —NHC(O)R⁹ and(C₁-C₈)alkanoyl; or R^(6a) and R^(6b) together form a spirocycle havingFormula (a); each R⁸ is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,(C₂-C₄)alkenyl, (C₂-C₄)alkynyl, aryl, heteroaryl, heterocycloalkyl orcycloalkyl, wherein R⁸ is optionally substituted with —OR, —N(R⁹)₂,—CON(R⁹)₂, or cyano; each R⁹ is independently H, (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, (C₂-C₄)alkenyl or (C₂-C₄)alkynyl; each n isindependently 0, 1 or 2; and m is 1, 2, 3, 4 or
 5. 2. The compound ofclaim 1, wherein A¹ is ethenylene, propenylene, butenylene, ethylene,propylene, butylene, oxypropylene, oxypropenylene, pyrazolylene,phenylene or pyrimidinylene.
 3. The compound of claim 1, wherein A² is—CH(R⁸)-quinolinylene, —CH(R⁸)-isoquinolinylene,—CH(R⁸)-naphthyridinylene, —CH(R⁸)-cinnolinylene,—CH(R⁸)-quinoxalinylene, —CH(R⁸)-phenylene or —CH(R⁸)-halophenylene. 4.The compound of claim 1, wherein R⁸ is methyl.
 5. The compound of claim1, wherein X¹ is —O—, —NH— or —N(CH₃)—; R¹ and R², when taken togetherwith the carbon to which they are both attached, form —C(═O)—; and R³ isH.
 6. The compound of claim 1, wherein X¹ is —O—, —NH— or —N(CH₃)—; R¹and R², when taken together with the carbon to which they are bothattached, form —C(═O)—; R³ is H; one of R^(4a) and R^(4b) is H and theother is methyl; and one of R^(5a) and R^(5b) is H and the other isisopropyl.
 7. The compound of claim 1, wherein X¹ is —O—, —NH— or—N(CH₃)—; R¹ and R², when taken together with the carbon to which theyare both attached, form —C(═O)—; R³ is H; one of R^(4a) and R^(4b) is Hand the other is methyl; one of R^(5a) and R^(5b) is H and the other isisopropyl; A² is —CH(R⁸)-heteroarylene; and R⁸ is methyl.
 8. Thecompound of claim 1, wherein one of R^(6a) and R^(6b) is H; and theother H, hydroxyl, —CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), (C₁-C₈)alkyl or(C₁-C₈)alkoxy, wherein R^(6a) or R^(6b) is optionally substituted withone or more substituents selected from the group consisting of halo,hydroxyl, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, heterocycloalkyl,(C₁-C₄)alkanoyl and di(C₁-C₄)alkylamino.
 9. The compound of claim 1,wherein R^(6a) and R^(6b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹.
 10. The compound of claim 1, which is a compound of FormulaII:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein: A¹ is

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)-cycloalkylene, arylene orcycloalkylene; X¹ is a bond, —CH₂—, —O—, —NH—, —N(CH₃)—,

R^(4a) is H, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, hydroxyl(C₁-C₈)alkyl,(C₁-C₄)alkoxy(C₁-C₈)alkyl or (C₁-C₈)alkyl; R⁵ is H, (C₁-C₄)alkyl,halo(C₁-C₈)alkyl, hydroxy(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₈)alkyl,cycloalkyl or heterocycloalkyl; R^(6a) and R^(6b) are independently H,hydroxyl, (C₁-C₈)alkyl, (C₂-C₈)alkenyl, (C₂-C₈)alkynyl, (C₁-C₈)alkoxy,—CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl, heterocycloalkyl, cycloalkyl, —SR⁹,—S(O)R⁹, —S(O)₂R⁹ or —N(R⁹)₂, wherein each of R^(6a) and R^(6b) isoptionally substituted with one or more substituents selected from thegroup consisting of halo, hydroxyl, cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy, aryl, cycloalkyl, heterocycloalkyl,mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹, —NHC(O)R⁹ and(C₁-C₈)alkanoyl where such an optional substitution is chemicallyfeasible; or R^(6a) and R^(6b) together form a spirocycle having Formula(a); or R^(6a) and R^(6b) together form a spirocycle having Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹; R^(7a) and R^(7b) are independently H, —OH, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl; R⁸ is H or (C₁-C₄)alky; and each R⁹is independently H, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₂-C₄)alkenyl or(C₂-C₄)alkynyl.
 11. The compound of claim 1, wherein R^(4a) is methyl;R⁵ is isopropyl; R⁸ is methyl; and A² is


12. The compound of claim 10, wherein R^(4a) is methyl; R⁵ is isopropyl;R⁸ is methyl; and A² is


13. The compound of claim 1, which is a compound of Formula II-c:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein: R^(6a) andR^(6b) are independently H, hydroxyl, (C₁-C₈)alkyl, (C₂-C₈)alkenyl,(C₂-C₈)alkynyl, (C₁-C₈)alkoxy, —CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy), aryl,heterocycloalkyl, cycloalkyl, —SR⁹, —S(O)R⁹, —S(O)₂R⁹ or —N(R⁹)₂,wherein each of R^(6a) and R^(6b) is optionally substituted with one ormore substituents selected from the group consisting of halo, hydroxyl,cyano, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, aryl, cycloalkyl,heterocycloalkyl, mono(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, —NHS(O)R⁹,—NHC(O)R⁹ and (C₁-C₈)alkanoyl where such an optional substitution ischemically feasible; or R^(6a) and R^(6b) together form a spirocyclehaving Formula (a); or R^(6a) and R^(6b) together form a spirocyclehaving Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, ═N(C₁-C₄)alkoxy, halo, hydroxyl, —NH₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, (C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and—S(O)₂R⁹; and each R⁹ is independently H, (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, (C₂-C₄)alkenyl or (C₂-C₄)alkynyl.
 14. The compound ofclaim 13, wherein R^(6a) and R^(6b) are independently H, —OH,(C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, (C₁-C₈)alkyl, hydroxy(C₁-C₈)alkyl,halo(C₁-C₈)alkyl, (C₁-C₄)alkoxy(C₁-C₈)alkyl,halo(C₁-C₄)alkoxy(C₁-C₈)alkyl,


15. The compound of claim 13, wherein R^(6a) and R^(6b) together form


16. The compound of claim 13, wherein R^(6a) and R^(6b) together form


17. The compound of claim 1, which is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 18. A compound whichis:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 19. A compound whichis:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 20. A compound whichis:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 21. A compound whichis:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 22. The compound ofclaim 1, which is a compound of Formula III:

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof, wherein L¹ is—O—CH₂—, —N(CH₃)—C(O)—, —NH—CH₂—, —N(CH₃)—CH₂—, —NH—CH(CF₃)— or—NH—S(O)₂—; A¹ is

A² is —CH(R⁸)-arylene, —CH(R⁸)-heteroarylene,—CH(R⁸)-heterocycloalkylene, —CH(R⁸)-cycloalkylene, arylene orcycloalkylene; X¹ is —O—, —N(CH₃)— or —NH—; R^(4a) is H, (C₁-C₄)alkoxy,halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl; R⁵ is H or (C₁-C₄)alkyl; R^(6a) andR^(6b) are independently H, hydroxyl, —CH₂CH₂CR⁹(═N(C₁-C₄)alkoxy),(C₁-C₈)alkyl or (C₁-C₈)alkoxy, optionally substituted with one or moresubstituents selected from the group consisting of halo, hydroxyl,(C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy, heterocycloalkyl, (C₁-C₄)alkanoyl ordi(C₁-C₄)alkylamino; or R^(6a) and R^(6b) together form a spirocyclehaving Formula (a):

wherein one or more carbon ring atoms of Formula (a) is optionallyreplaced by a nitrogen, oxygen or sulfur atom, and wherein a ring atomof Formula (a) optionally has one or more substituents selected from thegroup consisting of oxo, halo, hydroxyl, —NH₂, (C₁-C₄)alkyl,(C₁-C₄)alkoxy, —OC(O)R⁹, —C(O)₂R⁹, and —S(O)₂R⁹; R^(7a) and R^(7b) areindependently H, —OH, (C₁-C₄)alkoxy, halo(C₁-C₄)alkoxy or (C₁-C₈)alkyl;and R⁸ is H or (C₁-C₄)alkyl.
 23. The compound of claim 22, wherein L¹ is—NH—CH₂—; R^(4a) is methyl; R⁵ is isopropyl; R⁸ is methyl; and A² is


24. The compound of claim 23, wherein L¹ is —NH—S(O)₂—; R^(4a) ismethyl; R⁵ is isopropyl; R⁸ is methyl; and A² is


25. The compound of claim 23, wherein L¹ is —N(CH₃)—C(O)—; R^(4a) ismethyl; R⁵ is isopropyl; R⁸ is methyl; and A² is


26. The compound of claim 23, wherein L¹ is —O—CH₂—; R^(4a) is methyl;R⁵ is isopropyl; R⁸ is methyl; and A² is


27. The compound of claim 23, which is

or a pharmaceutically acceptable salt, isotope, stereoisomer, mixture ofstereoisomers, tautomer, ester or prodrug thereof.
 28. A pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof claim 1 or a pharmaceutically acceptable salt, isotope, stereoisomer,mixture of stereoisomers, tautomer, ester or prodrug thereof and apharmaceutically acceptable excipient.
 29. The pharmaceuticalcomposition of claim 28, further comprising at least one additionaltherapeutic agent selected from the group consisting of interferons,ribavirin, HCV NS3 protease inhibitors, HCV NS5a inhibitors, nucleosideor nucleotide inhibitors of HCV NS5B polymerase, non-nucleosideinhibitors of HCV NS5B polymerase, and TLR-7 agonists; or a mixturethereof.
 30. The pharmaceutical composition of claim 29, wherein the atleast one addition therapeutic agent is ribavirin, telaprevir,boceprevir or sofosbuvir.
 31. A method for treating a Flaviviridae viralinfection in a human patient in need thereof, comprising administeringto the patient a therapeutically effective amount of a compound of claim1 or a pharmaceutically acceptable salt, isotope, stereoisomer, mixtureof stereoisomers, tautomer, ester or prodrug thereof.
 32. The method ofclaim 31, wherein the viral infection is caused by a Hepatitis B virusor a Hepatitis C virus.
 33. The method of claim 32, wherein the viralinfection causes a disease selected from the group consisting of denguefever, yellow fever, hepatitis C, Japanese encephalitis, Kyasanur forestdisease, Murray valley encephalitis, St. Louis encephalitis, tick-borneencephalitis or West Nile encephalitis.
 34. A method for treating aCoronaviridae viral infection in a human patient in need thereof,comprising administering to the patient a therapeutically effectiveamount of a compound of claim 1 or a pharmaceutically acceptable salt,isotope, stereoisomer, mixture of stereoisomers, tautomer, ester orprodrug thereof.
 35. The method of claim 34, wherein the viral infectionis caused by a SARS coronarirus.
 36. The method of claim 35, wherein theviral infection causes a disease selected from the group consisting ofsevere acute respiratory syndrome (SARS), cancer, inflammation, obesity,acquired immune deficiency syndrome (AIDS), or cirrhosis.
 37. A methodfor providing immunomodulation to a human patient in need thereof,comprising administering to the patient a therapeutically effectiveamount of a compound of claim 1 or a pharmaceutically acceptable salt,isotope, stereoisomer, mixture of stereoisomers, tautomer, ester orprodrug thereof.